CN114532261B - Semi-viscous fish egg debonding agent and application thereof in artificial propagation of sinocyclocheilus grahami - Google Patents
Semi-viscous fish egg debonding agent and application thereof in artificial propagation of sinocyclocheilus grahami Download PDFInfo
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- CN114532261B CN114532261B CN202210115576.7A CN202210115576A CN114532261B CN 114532261 B CN114532261 B CN 114532261B CN 202210115576 A CN202210115576 A CN 202210115576A CN 114532261 B CN114532261 B CN 114532261B
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- 241000251468 Actinopterygii Species 0.000 title claims abstract description 77
- 241001275872 Sinocyclocheilus grahami Species 0.000 title claims abstract description 39
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 claims abstract description 38
- 229920001864 tannin Polymers 0.000 claims abstract description 35
- 239000001648 tannin Substances 0.000 claims abstract description 35
- 235000018553 tannin Nutrition 0.000 claims abstract description 35
- 229920001577 copolymer Polymers 0.000 claims abstract description 20
- 235000013601 eggs Nutrition 0.000 claims description 195
- 239000003795 chemical substances by application Substances 0.000 claims description 87
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 69
- 230000012447 hatching Effects 0.000 claims description 47
- 238000000034 method Methods 0.000 claims description 39
- 238000011534 incubation Methods 0.000 claims description 38
- 238000003756 stirring Methods 0.000 claims description 27
- 239000002202 Polyethylene glycol Substances 0.000 claims description 10
- 229920001223 polyethylene glycol Polymers 0.000 claims description 10
- 229920001442 polyethylene glycol-block-polycaprolactone Polymers 0.000 claims description 8
- BGGHCRNCRWQABU-JTQLQIEISA-N (2s)-2-amino-5-oxo-5-phenylmethoxypentanoic acid Chemical compound OC(=O)[C@@H](N)CCC(=O)OCC1=CC=CC=C1 BGGHCRNCRWQABU-JTQLQIEISA-N 0.000 claims description 5
- 108700004554 polyethylenimine-poly(gamma-benzylglutamate) Proteins 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 2
- 239000012528 membrane Substances 0.000 abstract description 15
- 241000894006 Bacteria Species 0.000 abstract description 5
- 241000233866 Fungi Species 0.000 abstract description 4
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 abstract description 3
- 239000001263 FEMA 3042 Substances 0.000 abstract description 3
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 abstract description 3
- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 abstract description 3
- 229940033123 tannic acid Drugs 0.000 abstract description 3
- 235000015523 tannic acid Nutrition 0.000 abstract description 3
- 229920002258 tannic acid Polymers 0.000 abstract description 3
- 230000001988 toxicity Effects 0.000 abstract description 3
- 231100000419 toxicity Toxicity 0.000 abstract description 3
- 238000013329 compounding Methods 0.000 abstract description 2
- 238000001179 sorption measurement Methods 0.000 abstract 1
- 238000005303 weighing Methods 0.000 description 82
- 238000005406 washing Methods 0.000 description 36
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 12
- 230000003203 everyday effect Effects 0.000 description 12
- 239000001301 oxygen Substances 0.000 description 12
- 229910052760 oxygen Inorganic materials 0.000 description 12
- 230000018109 developmental process Effects 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 239000000203 mixture Substances 0.000 description 5
- 239000000853 adhesive Substances 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000008144 egg development Effects 0.000 description 3
- 230000004720 fertilization Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 206010017533 Fungal infection Diseases 0.000 description 2
- 208000031888 Mycoses Diseases 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 230000001070 adhesive effect Effects 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000013020 embryo development Effects 0.000 description 2
- 230000009931 harmful effect Effects 0.000 description 2
- 239000001257 hydrogen Chemical group 0.000 description 2
- 229910052739 hydrogen Chemical group 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- HCWCAKKEBCNQJP-UHFFFAOYSA-N magnesium orthosilicate Chemical compound [Mg+2].[Mg+2].[O-][Si]([O-])([O-])[O-] HCWCAKKEBCNQJP-UHFFFAOYSA-N 0.000 description 2
- 239000000391 magnesium silicate Substances 0.000 description 2
- 229910052919 magnesium silicate Inorganic materials 0.000 description 2
- 235000019792 magnesium silicate Nutrition 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000013824 polyphenols Nutrition 0.000 description 2
- NGDUODKDUZDMOI-UHFFFAOYSA-M sodium;urea;chloride Chemical compound [Na+].[Cl-].NC(N)=O NGDUODKDUZDMOI-UHFFFAOYSA-M 0.000 description 2
- 239000000454 talc Substances 0.000 description 2
- 229910052623 talc Inorganic materials 0.000 description 2
- 235000012222 talc Nutrition 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000192497 Oscillatoria Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000012790 adhesive layer Substances 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000009027 insemination Effects 0.000 description 1
- YOBAEOGBNPPUQV-UHFFFAOYSA-N iron;trihydrate Chemical compound O.O.O.[Fe].[Fe] YOBAEOGBNPPUQV-UHFFFAOYSA-N 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000011345 viscous material Substances 0.000 description 1
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
Landscapes
- 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 discloses a semi-viscous fish egg debonder and application thereof in artificial propagation of sinocyclocheilus grahami, wherein the semi-viscous fish egg debonder comprises the following components in parts by weight: 20-40 parts of talcum powder, 1-3 parts of tannin and 5-10 parts of amphiphilic hyperbranched copolymer. By compounding the talcum powder, the tannin and the amphiphilic hyperbranched copolymer in a specific proportion, the adsorption of the talcum powder on the egg membrane of the fish egg is reduced, the toxicity of tannic acid on the egg membrane is reduced, the risk of bacteria and fungi infecting the egg membrane is reduced, and the hatchability of the sinocyclocheilus grahami egg is improved.
Description
Technical Field
The invention belongs to the field of aquaculture, and particularly relates to a semi-viscous fish egg debonder and application thereof in artificial propagation of sinocyclocheilus grahami.
Background
The fish producing viscous eggs or semi-viscous eggs have the problems of low fertility rate and hatchability in the artificial insemination and hatching process due to the viscosity of the fish eggs. For example, the egg membrane of fishes laying adhesive eggs or semi-adhesive eggs expands into a full and elastic adhesive layer when meeting water, and the fish eggs are attached to a substrate or a fish nest to cause the fish eggs to lose fertilization capability in a short time, so that the fertilization rate is low; moreover, fertilized eggs obtained after fertilization of the viscous eggs or semi-viscous eggs are easy to stick to the connecting blocks, so that internal gas exchange is difficult, and further the hatching rate is reduced. Therefore, it is necessary to detackify the viscous eggs or semi-viscous eggs.
The detackification method is a technique of inseminating after detackifying and then hatching in running water on the basis of artificial induced spawning. The debonding method is mainly divided into a physical debonding method and a chemical debonding method.
The physical debonding method mainly uses yellow mud, talcum powder, starch or Fuller's earth as debonding agent to remove the adhesive substance on the surface of fish egg under the action of mechanical shock and friction, so as to remove the adhesiveness of fish egg. The physical debonding method has loose requirements on the debonding treatment time, and the physical debonding agent does not generate harmful substances. However, the physical debonding method is easy to damage the fertilized eggs due to improper operation, and usually causes massive growth of bacteria and fungi during hatching, resulting in a decrease in hatching rate; in addition, the detackifier such as slurry and talcum powder is easy to form a layer of fine mud on the egg membrane of the fish egg, which is not beneficial to observing the embryo development.
The chemical detackification method is to remove the stickiness of the roe by treating it with a substance that chemically acts on the egg membrane of the roe, such as proteolytic enzyme, urea-sodium chloride mixed solution, tannin, or the like. The use of proteolytic enzymes, urea-sodium chloride mixed solutions or tannins can reduce the risk of bacterial and fungal infections. However, when the chemical debonding method is used to treat the fish eggs, the debonding operation needs to be completed in a short time, the dosage of the debonding agent is strictly required, and the improper dosage and the excessively long treatment time increase the mortality rate of the fertilized eggs, resulting in a decrease in the hatching rate.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a semi-viscous fish egg debonder and apply the debonder to the artificial propagation of sinocyclocheilus grahami. Through compounding specific proportion's talcum powder, tannin and amphiphilic hyperbranched copolymer, reduce the absorption of talcum powder on the egg membrane of roe, reduce the toxicity of tannic acid to the egg membrane, reduce the risk that bacterium and fungi infect the egg membrane, improve the hatchability of Yunnan oscillary roe.
The semi-viscous fish egg debonding agent comprises the following components in parts by weight: 20-40 parts of talcum powder, 1-3 parts of tannin and 5-10 parts of amphiphilic hyperbranched copolymer.
The talc contains hydrous magnesium silicate of talc as main ingredient and has a molecular formula of Mg 3 [Si 4 O 10 ](OH) 2 The compound is in a monoclinic system and has a layered structure, and the compound is used as a physical debonder to be applied to artificial propagation of sinocyclocheilus grahami, so that the fish egg can be effectively debonded.
Tannin is a natural high molecular compound with abundant resources, low price and easy obtainment, widely exists in plants, and is easily dissolved in solvents such as water, ethanol, acetone and the like. Tannins contain various reactive functional groups (phenolic hydroxyl, carboxyl) and show active chemical properties. Can react with the viscous substance on the fish egg membrane, thereby realizing the effect of debonding.
The amphiphilic hyperbranched copolymer contains a large number of hydroxyl groups and branched chains, the branched chains in the amphiphilic hyperbranched copolymer penetrate through the layered structure of the talcum powder, and hydrogen bonding effect exists between the branched chains and the hydrous magnesium silicate serving as the main component in the talcum powder, so that the adhesion amount of the talcum powder on an egg membrane of a fish egg is reduced, and the observation of embryo development is facilitated; hydroxyl in the amphiphilic hyperbranched copolymer and phenolic hydroxyl and carboxyl in tannin have hydrogen bond effects, reduce the toxicity of tannic acid to the egg membrane, effectively and properly prolong the debonding operation time, reduce the debonding operation difficulty, reduce the risk that the talcum powder incurs bacteria and fungi to infect the egg membrane during incubation, improve the hatchability of the sinocyclocheilus grahami roe.
Through the adaptation of talcum powder debonding agent and tannin debonding agent, the use of the reducible tannin debonding agent of the addition of talcum powder reduces the harm of tannin debonding agent to the egg membrane, and the risk that bacterium and fungal infection egg membrane are incurred to the talcum powder during the incubation in the addition of tannin debonding agent to improve the hatchability of Yunnan oscillatoria gracilis roe.
Preferably, the semi-viscous fish egg debonder comprises the following components in parts by weight: 25-35 parts of talcum powder, 1.5-2.5 parts of tannin and 6-9 parts of amphiphilic hyperbranched copolymer.
Preferably, the semi-viscous fish egg debonding agent comprises the following components in parts by weight: 25-30 parts of talcum powder, 1.5-2 parts of tannin and 7-8 parts of amphiphilic hyperbranched copolymer.
Preferably, the amphiphilic hyperbranched copolymer is selected from one or more of amphiphilic hyperbranched PEG-b-PCL, amphiphilic hyperbranched HBPO-g-DMA, amphiphilic hyperbranched PEI-PBLG, amphiphilic hyperbranched PEG-b-PEEGE, amphiphilic hyperbranched poly (L-glutamic acid-gamma-benzyl ester) -g-polyethylene glycol and amphiphilic hyperbranched polyaryletherketone-g-polyethylene glycol.
More preferably, the amphiphilic hyperbranched copolymer is selected from one or more of amphiphilic hyperbranched HBPO-g-DMA, amphiphilic hyperbranched poly (L-glutamic acid-gamma-benzyl ester) -g-polyethylene glycol and amphiphilic hyperbranched polyaryletherketone-g-polyethylene glycol.
Further, the amphiphilic hyperbranched copolymer also comprises 500-1000 parts by mass of water.
The invention also aims to provide a preparation method of the semi-viscous fish egg debonding agent, which comprises the following steps:
mixing pulvis Talci sieved by bolting silk, tannin, and amphiphilic hyperbranched copolymer, adding water, stirring well to obtain semi-viscous fish egg debonding agent.
The invention further aims to provide application of the semi-viscous fish egg debonder in artificial propagation of sinocyclocheilus grahami.
Further, the application of the semi-viscous fish egg debonder in the artificial propagation of the sinocyclocheilus grahami is to place fertilized eggs of the sinocyclocheilus grahami in the debonder for debonding and then hatching.
Further, the operation of hatching after debonding and debonding is to pour the debonding agent into the fertilized eggs, stir, wash off the debonding agent after debonding is completed, and hatch.
Preferably, the de-binding agitation time is 1-3 minutes. The debonding time of the debonding agent prepared by the scheme is moderate, and the debonding operation needs to be completed within 10S when the debonding agent is simply added with tannin, otherwise, harmful effects are generated, and the operation difficulty is reduced; meanwhile, the problems of long operation time and low debonding efficiency caused by the addition of a talcum powder debonding agent are avoided.
Preferably, the incubation temperature is 18-25 ℃ and the incubation pH is 6.5-7.5.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the following will clearly and completely describe the technical solution of the present invention with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, shall fall within the protection scope of the present invention.
Amphiphilic hyperbranched copolymers were purchased from sienna millennium biotechnology limited.
Example 1: and (4) preparing a debonding agent.
Weighing 20 parts by weight of talcum powder, 3 parts by weight of tannin and 10 parts by weight of amphiphilic hyperbranched PEG-b-PCL which are sieved by using 80-mesh bolting silk, adding into a beaker, adding 767 parts by weight of water, and uniformly stirring at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 2: and (4) preparing a debonding agent.
Weighing 40 parts by weight of talcum powder, 1 part by weight of tannin and 5 parts by weight of amphiphilic hyperbranched PEI-PBLG which are sieved by using 80-mesh bolting silk, adding into a beaker, adding 754 parts by weight of water, and stirring uniformly at room temperature to prepare the semi-viscous roe detackifier.
Example 3: and (3) preparing a debonding agent.
Weighing 25 parts by weight of talcum powder sieved by a 80-mesh bolting silk, 2.5 parts by weight of tannin, 9 parts by weight of amphiphilic hyperbranched PEG-b-PEEGE, adding into a beaker, adding 763.5 parts by weight of water, and stirring uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 4: and (4) preparing a debonding agent.
Weighing 35 parts by weight of talcum powder, 1.5 parts by weight of tannin and 6 parts by weight of amphiphilic hyperbranched PEG-b-PCL which are sieved and filtered by using 80-mesh bolting silk, adding the talcum powder, the tannin and the amphiphilic hyperbranched PEG-b-PCL into a beaker, adding 757.5 parts by weight of water, and uniformly stirring at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 5: and (4) preparing a debonding agent.
Weighing 30 parts by weight of talcum powder, 2 parts by weight of tannin and 7 parts by weight of amphiphilic hyperbranched PEI-PBLG which are sieved by using 80-mesh bolting silk, adding into a beaker, adding 761 parts by weight of water, and uniformly stirring at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 6: and (4) preparing a debonding agent.
30 parts by weight of talcum powder, 2 parts by weight of tannin and 8 parts by weight of amphiphilic hyperbranched PEG-b-PEEGE which are sieved and filtered by a 80-mesh bolting silk are weighed and added into a beaker, 760 parts by weight of water is added, and the mixture is stirred uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 7: and (4) preparing a debonding agent.
28 parts by weight of talcum powder, 1.8 parts by weight of tannin and 7.5 parts by weight of amphiphilic hyperbranched PEG-b-PCL which are sieved by a 80-mesh bolting silk are weighed and added into a beaker, 762.7 parts by weight of water are added, and the mixture is stirred uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 8: and (4) preparing a debonding agent.
28 parts by weight of talcum powder, 1.8 parts by weight of tannin and 7.5 parts by weight of amphiphilic hyperbranched HBPO-g-DMA which are sieved by using 80-mesh bolting silk are weighed and added into a beaker, 762.7 parts by weight of water are added, and the mixture is stirred uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 9: and (4) preparing a debonding agent.
28 parts by weight of talcum powder, 1.8 parts by weight of tannin and 7.5 parts by weight of amphiphilic hyperbranched poly (L-glutamic acid-gamma-benzyl ester) -g-polyethylene glycol which are sieved by a 80-mesh bolting silk are weighed and added into a beaker, 762.7 parts by weight of water are added, and the mixture is stirred uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 10: and (4) preparing a debonding agent.
28 parts by weight of talcum powder, 1.8 parts by weight of tannin and 7.5 parts by weight of amphiphilic hyperbranched polyaryletherketone-g-polyethylene glycol which are sieved by using 80-mesh bolting silk are weighed and added into a beaker, 762.7 parts by weight of water are added for dissolving, and the mixture is stirred uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
Example 11: and (3) debonding and incubating fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, putting fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 1 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing off the debonding agent prepared in the embodiment 1, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatching rate is 80.7%.
Example 12: and (3) carrying out debonding and incubation on fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, putting fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 2 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing the debonding agent prepared in the embodiment 2, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 80.1%.
Example 13: and (3) debonding and incubating fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, placing fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 3 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing the debonding agent prepared in the embodiment 3, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatching rate was 83.7%.
Example 14: and (3) debonding and incubating fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, placing fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
And after weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively and slowly pouring 10L of the debonding agent prepared in the embodiment 4 into a beaker along the inner wall of the container, stirring for 1-3 min, changing and washing for several times with clear water, washing away the debonding agent prepared in the embodiment 4, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5 during incubation, and observing and recording the fish egg development condition every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 84.2%.
Example 15: and (3) debonding and incubating fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, placing fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 5 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing off the debonding agent prepared in the embodiment 5, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 87.8%.
Example 16: and (3) carrying out debonding and incubation on fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, placing fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 6 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing the debonding agent prepared in the embodiment 6, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatching rate is 87.5 percent
Example 17: and (3) debonding and incubating fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, putting fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 7 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing the debonding agent prepared in the embodiment 7, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 86.9%.
Example 18: and (3) debonding and incubating fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, putting fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
And after weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively and slowly pouring 10L of the debonding agent prepared in the embodiment 8 into a beaker along the inner wall of the container, stirring for 1-3 min, changing and washing for several times with clear water, washing away the debonding agent prepared in the embodiment 8, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5 during incubation, and observing and recording the fish egg development condition every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 92.7%.
Example 19: and (3) carrying out debonding and incubation on fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, placing fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 9 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing the debonding agent prepared in the embodiment 9, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 92.6%.
Example 20: and (3) debonding and incubating fertilized eggs of the sinocyclocheilus grahami.
The method comprises the following steps of counting by adopting a weighing method, putting fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
And after weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively and slowly pouring 10L of the debonding agent prepared in the embodiment 10 into a beaker along the inner wall of a container, stirring for 1-3 min, changing and washing for several times with clear water, washing away the debonding agent prepared in the embodiment 10, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5 during incubation, and observing and recording the fish egg development condition every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 93.5%.
Comparative example 1
The method comprises the following steps of counting by adopting a weighing method, putting fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 1 into a beaker along the inner wall of a container, stirring for 30S, changing and washing for several times with clear water, washing off the debonding agent prepared in the embodiment 1, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5 during incubation, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatching rate was 62.4%.
Comparative example 2
The method comprises the following steps of counting by adopting a weighing method, placing fertilized eggs of the sinocyclocheilus grahami in a beaker, weighing 1kg of the fertilized eggs on an electronic weighing scale, weighing 2g of the fertilized eggs after weighing, counting, finally converting into the total number of the fertilized eggs, and recording the number of the fertilized eggs.
Weighing fertilized eggs, repeatedly washing the eggs with clear water for 2-3 times, adapting for 8min-10min, respectively slowly pouring 10L of the debonding agent prepared in the embodiment 1 into a beaker along the inner wall of a container, stirring for 4 min, changing and washing for several times with clear water, washing off the debonding agent prepared in the embodiment 1, pouring the debonding agent into an incubator for running water incubation, wherein the water temperature is 18-25 ℃, the dissolved oxygen is 6mg/L-8mg/L, the incubation pH is 6.5-7.5, and observing and recording the development condition of the fish eggs every day. And (3) calculating the hatching rate, wherein the hatching rate calculation formula is as follows: hatchability = number of hatched fish larvae/fertilized eggs × 100%. The hatchability was 68.7%.
Comparative example 3
Weighing 20 parts by weight of talcum powder sieved by a 80-mesh bolting silk, adding the talcum powder into a beaker, adding 780 parts by weight of water, and stirring uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
The prepared semi-viscous fish egg debonding agent is applied to debonding and hatching of the fertilized egg of sinocyclocheilus grahami, and the steps of the debonding and hatching are the same as those in the embodiment 12. The hatchability was 35.7%.
Comparative example 4
Weighing 3 parts by weight of tannin sieved by a 80-mesh bolting silk into a beaker, adding 797 parts by weight of water, and stirring uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
The prepared semi-viscous fish egg debonding agent is applied to debonding and hatching of the fertilized egg of sinocyclocheilus grahami, and the steps of the debonding and hatching are the same as those in the embodiment 12. The hatchability was 41.6%.
Comparative example 5
Weighing 20 parts by weight of talcum powder and 3 parts by weight of tannin which are sieved and filtered by a 80-mesh bolting silk into a beaker, adding 777 parts by weight of water, and stirring uniformly at room temperature to prepare the semi-viscous fish egg debonding agent.
The obtained semi-viscous fish egg debonder is applied to the debonding incubation of the fertilized egg of sinocyclocheilus grahami, and the steps of the debonding incubation are the same as the example 12. The hatching rate is 76.8%.
Comparative example 6
Weighing 20 parts by weight of talcum powder sieved by using 80-mesh bolting silk and 10 parts by weight of amphiphilic hyperbranched PEG-b-PCL, adding into a beaker, adding 770 parts by weight of water, and uniformly stirring at room temperature to prepare the semi-viscous fish egg debonding agent.
The prepared semi-viscous fish egg debonding agent is applied to debonding and hatching of the fertilized egg of sinocyclocheilus grahami, and the steps of the debonding and hatching are the same as those in the embodiment 12. The hatching rate was 42.6%.
Comparative example 7
Weighing 3 parts by weight of tannin sieved by a 80-mesh bolting silk and 10 parts by weight of amphiphilic hyperbranched PEG-b-PCL, adding into a beaker, adding 787 parts by weight of water for dissolving, and stirring uniformly at room temperature to prepare the semi-viscous roe debonding agent.
The prepared semi-viscous fish egg debonding agent is applied to debonding and hatching of the fertilized egg of sinocyclocheilus grahami, and the steps of the debonding and hatching are the same as those in the embodiment 12. The hatchability was 65.4%.
Finally, it should be noted that the above-mentioned embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the same, and although the present invention is described in detail with reference to the above-mentioned embodiments, it should be understood by those skilled in the art that after reading the present specification, modifications and equivalents of the specific embodiments of the present invention can be made, but these modifications and variations do not depart from the scope of the claims of the present application.
Claims (9)
1. The semi-viscous fish egg debonding agent is characterized by comprising the following components in parts by weight: 20-40 parts of talcum powder, 1-3 parts of tannin and 5-10 parts of amphiphilic hyperbranched copolymer; the amphiphilic hyperbranched copolymer is selected from one or more of amphiphilic hyperbranched PEG-b-PCL, amphiphilic hyperbranched HBPO-g-DMA, amphiphilic hyperbranched PEI-PBLG, amphiphilic hyperbranched PEG-b-PEEGE, amphiphilic hyperbranched poly (L-glutamic acid-gamma-benzyl ester) -g-polyethylene glycol and amphiphilic hyperbranched polyaryletherketone-g-polyethylene glycol.
2. The semi-viscous fish egg debonder of claim 1, wherein the semi-viscous fish egg debonder comprises the following components in parts by weight: 25-35 parts of talcum powder, 1.5-2.5 parts of tannin and 6-9 parts of amphiphilic hyperbranched copolymer.
3. The semi-viscous fish egg debonder of claim 1, wherein the semi-viscous fish egg debonder comprises the following components in parts by weight: 25-30 parts of talcum powder, 1.5-2 parts of tannin and 7-8 parts of amphiphilic hyperbranched copolymer.
4. The semi-viscous fish egg debonder of claim 1 wherein the amphiphilic hyperbranched copolymer is selected from one or more of amphiphilic hyperbranched HBPO-g-DMA, amphiphilic hyperbranched poly (L-glutamic acid- γ -benzyl ester) -g-polyethylene glycol, amphiphilic hyperbranched polyaryletherketone-g-polyethylene glycol.
5. A method of preparing the semi-viscous egg detackifier of claim 1, comprising the steps of:
mixing talcum powder, tannin and amphiphilic hyperbranched copolymer which are sieved by bolting silk, adding water, and stirring uniformly to prepare the semi-viscous fish egg debonding agent.
6. Use of a semi-viscous roe detackifier according to any one of claims 1 to 3 in the artificial reproduction of sinocyclocheilus grahami.
7. A method of using the semi-viscous egg detackifier of claims 1-4, comprising: and (4) putting the fertilized eggs into a debonding agent for debonding and then hatching.
8. The method for using the semi-viscous fish egg debonding agent according to claim 7, wherein the post-debonding incubation operation is to pour the debonding agent into the fertilized eggs, stir, wash off the debonding agent after the debonding is completed, and incubate.
9. The method of using a semi-viscous egg debonder according to claim 7, wherein the debonding stirring time is 1 to 3 minutes.
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