CN117084204B - Buffer solution for improving hatching rate and survival rate of eastern conch oocysts and method for hatching eastern conch oocysts - Google Patents
Buffer solution for improving hatching rate and survival rate of eastern conch oocysts and method for hatching eastern conch oocysts Download PDFInfo
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- CN117084204B CN117084204B CN202311273789.3A CN202311273789A CN117084204B CN 117084204 B CN117084204 B CN 117084204B CN 202311273789 A CN202311273789 A CN 202311273789A CN 117084204 B CN117084204 B CN 117084204B
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- oocysts
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- conch
- babylonia
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- 210000003250 oocyst Anatomy 0.000 title claims abstract description 105
- 230000012447 hatching Effects 0.000 title claims abstract description 96
- 230000004083 survival effect Effects 0.000 title claims abstract description 41
- 239000007853 buffer solution Substances 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 24
- 241000606434 Babylonia Species 0.000 claims abstract description 65
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 28
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims abstract description 28
- 239000013535 sea water Substances 0.000 claims abstract description 21
- AYIRNRDRBQJXIF-NXEZZACHSA-N (-)-Florfenicol Chemical compound CS(=O)(=O)C1=CC=C([C@@H](O)[C@@H](CF)NC(=O)C(Cl)Cl)C=C1 AYIRNRDRBQJXIF-NXEZZACHSA-N 0.000 claims abstract description 18
- QTDMGAWIBXJNRR-UHFFFAOYSA-N Mangostin Natural products CC(=CCc1c(O)cc2Oc3cc(C)c(O)c(CC=C(C)C)c3C(=O)c2c1O)C QTDMGAWIBXJNRR-UHFFFAOYSA-N 0.000 claims abstract description 18
- QNVSXXGDAPORNA-UHFFFAOYSA-N Resveratrol Natural products OC1=CC=CC(C=CC=2C=C(O)C(O)=CC=2)=C1 QNVSXXGDAPORNA-UHFFFAOYSA-N 0.000 claims abstract description 18
- LUKBXSAWLPMMSZ-OWOJBTEDSA-N Trans-resveratrol Chemical compound C1=CC(O)=CC=C1\C=C\C1=CC(O)=CC(O)=C1 LUKBXSAWLPMMSZ-OWOJBTEDSA-N 0.000 claims abstract description 18
- GNRIZKKCNOBBMO-UHFFFAOYSA-N alpha-mangostin Chemical compound OC1=C(CC=C(C)C)C(O)=C2C(=O)C3=C(CC=C(C)C)C(OC)=C(O)C=C3OC2=C1 GNRIZKKCNOBBMO-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229960003760 florfenicol Drugs 0.000 claims abstract description 18
- 235000021283 resveratrol Nutrition 0.000 claims abstract description 18
- 229940016667 resveratrol Drugs 0.000 claims abstract description 18
- ZVFQDLCERPGZMO-UHFFFAOYSA-N alpha-mangostin Natural products OC1=C(CC=C(C)C)C(O)=C2C(=O)C3=C(CC=C(C)C)C(OC)=C(O)C=C3CC2=C1 ZVFQDLCERPGZMO-UHFFFAOYSA-N 0.000 claims abstract description 17
- PMYDPQQPEAYXKD-UHFFFAOYSA-N 3-hydroxy-n-naphthalen-2-ylnaphthalene-2-carboxamide Chemical compound C1=CC=CC2=CC(NC(=O)C3=CC4=CC=CC=C4C=C3O)=CC=C21 PMYDPQQPEAYXKD-UHFFFAOYSA-N 0.000 claims abstract description 14
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims abstract description 14
- PMVSDNDAUGGCCE-TYYBGVCCSA-L Ferrous fumarate Chemical compound [Fe+2].[O-]C(=O)\C=C\C([O-])=O PMVSDNDAUGGCCE-TYYBGVCCSA-L 0.000 claims abstract description 14
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 14
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims abstract description 14
- DFPOZTRSOAQFIK-UHFFFAOYSA-N S,S-dimethyl-beta-propiothetin Chemical compound C[S+](C)CCC([O-])=O DFPOZTRSOAQFIK-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229930003268 Vitamin C Natural products 0.000 claims abstract description 14
- FNAQSUUGMSOBHW-UHFFFAOYSA-H calcium citrate Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FNAQSUUGMSOBHW-UHFFFAOYSA-H 0.000 claims abstract description 14
- 239000001354 calcium citrate Substances 0.000 claims abstract description 14
- 235000002332 ferrous fumarate Nutrition 0.000 claims abstract description 14
- 239000011773 ferrous fumarate Substances 0.000 claims abstract description 14
- 229960000225 ferrous fumarate Drugs 0.000 claims abstract description 14
- 235000001727 glucose Nutrition 0.000 claims abstract description 14
- 239000008103 glucose Substances 0.000 claims abstract description 14
- 229940004916 magnesium glycinate Drugs 0.000 claims abstract description 14
- AACACXATQSKRQG-UHFFFAOYSA-L magnesium;2-aminoacetate Chemical compound [Mg+2].NCC([O-])=O.NCC([O-])=O AACACXATQSKRQG-UHFFFAOYSA-L 0.000 claims abstract description 14
- 239000001103 potassium chloride Substances 0.000 claims abstract description 14
- 235000011164 potassium chloride Nutrition 0.000 claims abstract description 14
- 235000018716 sodium selenate Nutrition 0.000 claims abstract description 14
- 239000011655 sodium selenate Substances 0.000 claims abstract description 14
- 229960001881 sodium selenate Drugs 0.000 claims abstract description 14
- 235000013337 tricalcium citrate Nutrition 0.000 claims abstract description 14
- 235000019154 vitamin C Nutrition 0.000 claims abstract description 14
- 239000011718 vitamin C Substances 0.000 claims abstract description 14
- 239000011701 zinc Substances 0.000 claims abstract description 14
- 229910052725 zinc Inorganic materials 0.000 claims abstract description 14
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 150000001413 amino acids Chemical class 0.000 claims abstract description 11
- 229960001196 thiotepa Drugs 0.000 claims abstract description 10
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims abstract description 5
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 5
- 229960004256 calcium citrate Drugs 0.000 claims abstract description 5
- 229960001031 glucose Drugs 0.000 claims abstract description 5
- 229960002816 potassium chloride Drugs 0.000 claims abstract description 5
- 239000012528 membrane Substances 0.000 claims description 18
- 241000237858 Gastropoda Species 0.000 claims description 11
- 239000000872 buffer Substances 0.000 claims description 9
- 238000009395 breeding Methods 0.000 claims description 6
- 230000001488 breeding effect Effects 0.000 claims description 6
- 238000011534 incubation Methods 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 230000007480 spreading Effects 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- XBDQKXXYIPTUBI-UHFFFAOYSA-N Propionic acid Substances CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- -1 zinc amino acid Chemical class 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 229920006395 saturated elastomer Polymers 0.000 claims description 2
- ZAKOWWREFLAJOT-CEFNRUSXSA-N D-alpha-tocopherylacetate Chemical compound CC(=O)OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-CEFNRUSXSA-N 0.000 claims 1
- FOCVUCIESVLUNU-UHFFFAOYSA-N Thiotepa Chemical compound C1CN1P(N1CC1)(=S)N1CC1 FOCVUCIESVLUNU-UHFFFAOYSA-N 0.000 claims 1
- ZAKOWWREFLAJOT-UHFFFAOYSA-N d-alpha-Tocopheryl acetate Natural products CC(=O)OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-UHFFFAOYSA-N 0.000 claims 1
- 230000008901 benefit Effects 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 4
- 239000001963 growth medium Substances 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 20
- 235000013601 eggs Nutrition 0.000 description 10
- 230000007246 mechanism Effects 0.000 description 9
- OSVXSBDYLRYLIG-UHFFFAOYSA-N dioxidochlorine(.) Chemical compound O=Cl=O OSVXSBDYLRYLIG-UHFFFAOYSA-N 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 241000565675 Oncomelania Species 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 241001247197 Cephalocarida Species 0.000 description 3
- 239000004155 Chlorine dioxide Substances 0.000 description 3
- 235000019398 chlorine dioxide Nutrition 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 235000015170 shellfish Nutrition 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 208000035404 Autolysis Diseases 0.000 description 2
- 206010057248 Cell death Diseases 0.000 description 2
- 241000237852 Mollusca Species 0.000 description 2
- 229910000831 Steel Inorganic materials 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000029052 metamorphosis Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000003204 osmotic effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 230000028043 self proteolysis Effects 0.000 description 2
- 239000010959 steel Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 1
- 241000920062 Babylonia areolata Species 0.000 description 1
- 235000001815 DL-alpha-tocopherol Nutrition 0.000 description 1
- 239000011627 DL-alpha-tocopherol Substances 0.000 description 1
- 241000233805 Phoenix Species 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000002180 anti-stress Effects 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 210000000625 blastula Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 238000013100 final test Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000008182 oocyte development Effects 0.000 description 1
- HDMGAZBPFLDBCX-UHFFFAOYSA-M potassium;sulfooxy sulfate Chemical compound [K+].OS(=O)(=O)OOS([O-])(=O)=O HDMGAZBPFLDBCX-UHFFFAOYSA-M 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 229940042585 tocopherol acetate Drugs 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/51—Culture of aquatic animals of shellfish of gastropods, e.g. abalones or turban snails
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; 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
Abstract
The invention discloses a buffer solution for improving hatching rate and survival rate of Babylonia oocysts and a hatching method of Babylonia oocysts based on the buffer solution. The buffer solution comprises alpha-mangostin, resveratrol, florfenicol, dimethyl-beta-thiotepa DMPT, magnesium glycinate, potassium chloride, calcium citrate, vitamin C, DL-tocopheryl acetate, proline, an amino acid zinc complex, sodium selenate, ferrous fumarate, glucose and sterile seawater. The culture medium is specially used for hatching the eastern conch oocysts, solves the problem that the eastern conch oocysts cannot be hatched smoothly in the process of hatching, greatly improves the hatching rate and hatching speed of the eastern conch oocysts, ensures that the hatching rate of the eastern conch oocysts is more than or equal to 90%, ensures that the survival rate is more than or equal to 89%, and greatly improves the economic benefit of eastern conch seedling culture.
Description
Technical Field
The invention belongs to the technical field of shellfish seedling raising on aquatic products, and particularly relates to a buffer solution for improving hatching rate and survival rate of eastern conch oocysts and a method for hatching the eastern conch oocysts.
Background
Babylonia is commonly called as oncomelania, oncomelania, and is also called as phoenix in Taiwan, and belongs to the phylum Mollusca (Mollusca), gastropoda (Gastropoda), front gill subclass (Prosobranchia), new Abdominal order (Neopatropoda), bucciidae (Bucciidae), babylonia (Babylonia), and is distributed in tropical and subtropical areas, and the main breeding provinces of Babylonia in China are four provinces of Hainan, guangdong, fujian and Guangxi, and the breeding varieties mainly include Babylonia with square spots, babylonia and Babylonia. Babylonia is a meat shallow seafloor shellfish, has delicious taste, and is a high-protein low-fat rare seafood meeting the modern nutritional requirements.
Babylonia mainly depends on sea water fishing to cause serious decay of natural resources, and the Babylonia can not meet the market demand by relying on natural shellfish fishing for a long time. Therefore, only through the research of artificial breeding technology, the artificial breeding of a large number of breeding seeds can meet a large number of demands of the market on the Babylonia, and meanwhile, the natural resource protection of the Babylonia is also positively acted.
The artificial seedling raising technology for Babylonia with square spots includes such steps as egg collection, egg hatching, larva cultivation, bait kind and larva cultivation effect, and features low survival rate and metamorphosis rate of Babylonia larva, and wide variation range of metamorphosis rate.
In addition, in the process of raising the seedlings of Babylonia, the phenomenon that part of oocysts cannot hatch or hatch difficultly often occurs, and the hatching difficulty is sometimes as high as 50%. If only the eastern conch oocysts are subject to autolysis and hydrolysis, along with the prolongation of autolysis time, the larvae in the membrane and oocyst membranes can be infected, the larvae die in the membrane, and the pollution to the water body is also influenced. If the oocysts are broken, the complete extracapsular culture faces the problem of extracapsular implantation of the Babylonia larvae and the problem of extracapsular life support, namely the problem of how to simulate the internal environment of the Babylonia oocysts, the composition of endosperm, the requirements of different nutrients for different stages of development and vital activities, extracapsular development infection and the like.
Therefore, how to solve the problems of low hatching rate of the eastern conch oocysts and difficult hatching of larvae by self membrane rupture, explore a culture solution and a method for effectively and conveniently improving the hatching rate and the survival rate of the eastern conch, and become a technical problem to be solved urgently.
Disclosure of Invention
Aiming at the prior art problems, the primary aim of the invention is to provide the buffer solution for improving the hatching rate and the survival rate of the eastern conch oocysts, which is not only suitable for hatching the eastern conch oocysts, but also solves the problem that the eastern conch oocysts cannot be hatched smoothly in the process of hatching, greatly improves the hatching rate and the survival rate of the eastern conch oocysts, and improves the economic benefit of eastern conch seedling raising.
The second aim of the invention is to provide a preparation method of the buffer solution for improving the hatching rate and the survival rate of the eastern wind snail oocysts.
A third object of the present invention is to provide the use of the above-described buffer in hatching or increasing hatching rate and survival rate of eastern conch oocysts.
A fourth object of the present invention is to provide a method of hatching eastern oncomelania oocysts.
In order to achieve the above object, the present invention is realized by the following technical scheme:
a buffer solution for improving hatching rate and survival rate of eastern snail oocysts comprises 0.1-0.3 g/L alpha-mangostin, 0.1-0.3 g/L resveratrol, 0.02-0.03 g/L florfenicol, 0.2-0.9 g/L dimethyl-beta-propionic acid thiopavilion DMPT, 1.0-5.0 g/L magnesium glycinate, 0.5-1.0 g/L potassium chloride, 1.0-5.0 g/L calcium citrate, 0.03-0.06 g/L vitamin C, 0.02-0.06 g/L LDL-tocopheryl acetate, 0.02-0.04 g/L proline, 0.01-0.03 g/L amino acid zinc complex, 0.01-0.03 g/L sodium selenate, 0.01-0.03 g/L ferrous fumarate, 5.0-10.0 g/L glucose and 1L seawater for disinfection.
The invention provides a buffer solution for improving the hatching rate and the survival rate of the eastern conch oocysts, and the inventor discovers through researches,
the alpha-mangostin, the resveratrol and the florfenicol have good antibacterial and antioxidant effects and good adaptability to the larvae of the Babylonia, and under the cooperation of the alpha-mangostin, the resveratrol and the florfenicol, the hatching rate and the survival rate of the oocysts and the larvae of the Babylonia with difficult hatching can be greatly improved and promoted.
Further, the dimethyl-beta-thiotepa DMPT in the buffer solution is used as an in-vivo osmotic pressure buffer, so that the anti-stress capability and the anti-osmotic pressure effect of the mechanical membrane rupture can be improved;
the coordination of magnesium glycinate, potassium chloride, calcium citrate, vitamin C, LDL-tocopheryl acetate, proline, amino acid zinc complex, sodium selenate, ferrous fumarate and glucose not only can provide proper osmotic pressure for the Babylonia larvae, but also contains various nutrients which are beneficial to improving the immunity and the vitality of organisms, regulating metabolism and promoting growth and development.
Under the combined action, the adaptability and the survival rate of the Babylonia larvae after mechanical rupture of membranes are greatly improved, meanwhile, the infection of the Babylonia oocysts in the water body is reduced, and the economic benefit of seedling raising of the Babylonia is increased.
Preferably, the buffer solution comprises 0.2-0.3 g/L alpha-mangostin, 0.2-0.3 g/L resveratrol, 0.02-0.03 g/L florfenicol, 0.5-0.8 g/L dimethyl-beta-thiotepa DMPT, 3.0-4.0 g/L magnesium glycinate, 0.7-8.0 g/L potassium chloride, 4.0-5.0 g/L calcium citrate, 0.04-0.05 g/L vitamin C, 0.05-0.06 g/L DL-tocopherol acetate, 0.03-0.04 g/L proline, 0.02-0.03 g/L amino acid zinc complex, 0.02-0.03 g/L sodium selenate, 0.02-0.03 g/L ferrous fumarate, 6.0-8.0 g/L glucose, and 1L disinfection seawater.
Furthermore, the invention claims a preparation method of a buffer solution for improving the hatching rate and the survival rate of the eastern snail oocysts, which is characterized in that sterile seawater, alpha-mangostin, resveratrol, florfenicol, dimethyl-beta-propiothiazole DMPT, magnesium glycinate, potassium chloride, calcium citrate, vitamin C, DL-tocopheryl acetate, proline, an amino acid zinc complex, sodium selenate, ferrous fumarate and glucose are mixed and stirred uniformly to obtain the buffer solution for improving the hatching rate of the eastern snail oocysts.
Furthermore, the invention claims the application of the buffer solution in hatching of the eastern conch oocysts or improving the hatching rate and the survival rate of the eastern conch oocysts.
Further, the invention also claims a method for hatching the eastern conch oocysts, which comprises the following steps:
s1, uniformly spreading eastern conch oocysts at the bottom of an incubation frame, and puncturing the eastern conch oocysts until all larvae can break membranes by themselves; or the eastern conch oocysts are evenly spread at the bottom of the hatching frame, and the hatching frame is placed under the air pressure, so that the eastern conch oocyst membranes are crushed under the air pressure;
s2, stopping oxygen for 30 minutes after 10 minutes of inflation, fishing up floating Babylonia larvae, and discarding the Babylonia larvae at the bottom layer;
s3, cleaning the floating Babylonia larvae in the step S2, mixing and soaking the Babylonia larvae, the sterilized seawater and the buffer solution for improving the hatching rate of the Babylonia oocysts for 1-3 hours, and then pouring the mixed solution into a seedling raising pond for raising seedlings.
Preferably, the eastern conch oocysts are mature eastern conch oocysts, larvae inside the oocysts are developed into membranous internal disc larvae, the appearance of the oocysts is changed from light yellow to earthy yellow and finally to dark brown, the edges of the oocysts are changed into black, and endosperm liquid of the oocysts is changed from a viscous state to a water-like state to be in a saturated state.
Preferably, in the step S3, the concentration of the buffer solution for improving the hatching rate of the eastern wind snail oocysts is 3-10 ppm.
Preferably, the Babylonia is selected from one or more of Babylonia quadricarina, babylonia taiwan or Babylonia.
Preferably, in the step S3, the density of the eastern oncomelania larvae in the seedling raising pond is 60-120/L.
Preferably, in the step S3, the disinfection of the seawater is performed by using chlorine dioxide with a concentration of 2-4 ppm or potassium hydrogen persulfate with a concentration of 1-3 ppm.
Preferably, in the step S3, the volume of the sterilized seawater is equal to or greater than 50L.
Preferably, in the step S1, the air pressure ranges from 200 kPa to 400 kPa.
Preferably, in the step S1, the eastern conch oocysts may be punctured by a needle body with a spike-shaped end, or may be punctured by an egg-breaking cone. Preferably, the egg breaking cone consists of a handle, a mechanism main body, a supporting mechanism, a breaking thimble, a lifting mechanism, a button and a movable baffle plate. The supporting mechanisms are uniformly distributed at intervals, the installation direction of each supporting mechanism is perpendicular to the main body of the mechanism, 1 crushing thimble is arranged at the lower end of each supporting mechanism, a movable baffle is arranged in the middle of each crushing needle, a plurality of lifting mechanisms with uniform intervals are correspondingly arranged at the upper end of each movable baffle, each lifting mechanism comprises a lifting plate and a POM spring, and the springs are arranged between the movable baffle and the lifting plate and are connected with each other; the movable baffle plate is correspondingly provided with a thimble hole corresponding to the position of the crushing thimble, the crushing thimble on the lifting plate can correspondingly pass through the thimble hole, and the movable baffle plate can correspondingly retract upwards to the lifting plate and extend downwards to the tail end of the crushing thimble. When the oocysts are pricked, if the oocysts still remain on the steel needle, the button can be pressed lightly, the movable baffle can eject the residual oocysts out of the steel needle, and then the residual oocysts can automatically rebound to the original position.
Compared with the prior art, the invention has the following beneficial effects: the buffer solution is specially used for hatching the eastern conch oocysts, solves the problem that the eastern conch oocysts cannot be hatched smoothly in the process of hatching, greatly improves the hatching rate and hatching speed of the eastern conch oocysts, ensures that the hatching rate of the eastern conch oocysts is more than or equal to 90 percent, ensures that the survival rate is more than or equal to 89 percent, and greatly improves the economic benefit of eastern conch seedling raising.
Drawings
FIG. 1 is a schematic representation of the hatching difficulty of Babylonia areolata oocysts in example 5.
FIG. 2 is a schematic representation of the hatching of Babylonia larvae after puncturing the oocysts with needles in example 5.
FIG. 3 is a schematic representation of infection decay after continued incubation of Babylonia quadricarina oocysts of comparative example 3 without artificial rupture of the membranes.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Example 1 preparation of buffer solution for increasing hatching Rate and survival Rate of Dongfeng spiral oocysts
1L of sterilized seawater is prepared, 0.2g of alpha-mangostin, 0.2g of resveratrol, 0.03g of florfenicol, 0.5g of dimethyl-beta-thiotepa DMPT,4.0g of magnesium glycinate, 0.7g of potassium chloride, 4.0g of calcium citrate, 0.05g of vitamin C,0.05 gDL-tocopheryl acetate, 0.03g of proline, 0.02g of amino acid zinc complex, 0.02g of sodium selenate, 0.02g of ferrous fumarate and 8.0g of glucose are sequentially added, and the buffer solution for improving the hatching rate of the eastern-snail oocysts is prepared after uniform stirring.
Example 2 preparation of buffer solution for increasing hatching Rate and survival Rate of Dongfeng spiral oocysts
1L of sterilized seawater is prepared, 0.3g of alpha-mangostin, 0.3g of resveratrol, 0.02g of florfenicol, 0.8g of dimethyl-beta-thiotepa DMPT,3.0g of magnesium glycinate, 0.8g of potassium chloride, 4.5g of calcium citrate, 0.04g of vitamin C,0.06 gDL-tocopheryl acetate, 0.04g of proline, 0.03g of amino acid zinc complex, 0.03g of sodium selenate, 0.03g of ferrous fumarate and 6.0g of glucose are sequentially added, and after uniform stirring, the buffer solution for improving the hatching rate of the eastern-snail oocysts is prepared.
Example 3 preparation of buffer solution for increasing hatching Rate and survival Rate of Dongfeng spiral oocysts
1L of sterilized seawater is prepared, 0.1g of alpha-mangostin, 0.1g of resveratrol, 0.02g of florfenicol, 0.2g of dimethyl-beta-thiotepa DMPT,1.0g of magnesium glycinate, 0.5g of potassium chloride, 1g of calcium citrate, 0.03g of vitamin C,0.02 gDL-tocopheryl acetate, 0.02g of proline, 0.01g of amino acid zinc complex, 0.01g of sodium selenate, 0.01g of ferrous fumarate and 5.0g of glucose are sequentially added, and the buffer solution for improving the hatching rate of the eastern-snail oocysts is prepared after uniform stirring.
Example 4 preparation of buffer solution for increasing hatching Rate and survival Rate of Dongfeng spiral oocysts
1L of sterilized seawater is prepared, 0.3g of alpha-mangostin, 0.3g of resveratrol, 0.02g of florfenicol, 0.9g of dimethyl-beta-thiotepa DMPT,5.0g of magnesium glycinate, 1.0g of potassium chloride, 5.0g of calcium citrate, 0.06g of vitamin C,0.02 gDL-tocopheryl acetate, 0.02g of proline, 0.01g of amino acid zinc complex, 0.01g of sodium selenate, 0.01g of ferrous fumarate and 10.0g of glucose are sequentially added, and after uniform stirring, the buffer solution for improving the hatching rate of the eastern-snail oocysts is prepared.
Example 5 method of hatching Dongfeng spiral oocysts
(1) As shown in fig. 1, uniformly spreading the eastern Babylonia oocysts with difficult hatching at the bottom of a hatching frame, and repeatedly puncturing the eastern Babylonia oocysts by a needle until all larvae can break membranes by themselves (as shown in fig. 2);
(2) Stopping oxygen for 10 minutes after 3 minutes of inflation, fishing up floating Babylonia larvae by using a 60-mesh dip net, and discarding the Babylonia larvae at the bottom layer;
(3) The floating Babylonia larvae are washed by seawater disinfected by 2ppm of chlorine dioxide and then put into a 100L hatching artemia barrel filled with disinfected seawater, 6ppm of buffer solution prepared in example 1 is put into the barrel in advance, the whole basin is poured into a seedling raising pond after 2 hours of soaking bath, the density of the larvae in the seedling raising pond is kept to be 70-80 per liter, the hatching rate is 92%, and the survival rate is 93.5%.
Example 6 method of hatching Dongfeng spiral oocysts
(1) Uniformly spreading the hatching difficult Babylonia oocysts at the bottom of a hatching frame, and placing the hatching frame under an air pressure controller (a VPC2021 series controller manufactured by Yiyang company, the air pressure is regulated to be 300 kPa), so that the Babylonia oocyst membranes are crushed under the air pressure;
(2) Stopping oxygen for 8 minutes after 2 minutes of inflation, fishing up floating Babylonia larvae by using a 60-mesh dip net, and discarding the Babylonia larvae at the bottom layer;
(3) The floating Babylonia larvae are washed by seawater disinfected by 2ppm of chlorine dioxide and then put into a 100L hatching artemia barrel filled with disinfected seawater, 6ppm of buffer solution prepared in example 1 is put into the barrel in advance, the whole basin is poured into a seedling raising pond after 2 hours of soaking bath, and the density of the larvae in the seedling raising pond is kept to be 70-80 per liter, the hatching rate is 90%, and the survival rate is 89%.
Comparative example 1
The difference between this comparative example and example 5 is that: step (1) and step (2) are not carried out, and buffer solution is not adopted to carry out bath on the Babylonia larvae; the specific operation is as follows: uniformly spreading the eastern Babylonia oocysts which are difficult to hatch at the bottom of a hatching frame, putting the eastern Babylonia oocysts into a 100L hatching artemia barrel filled with sterile seawater for hatching, and then transferring the eastern Babylonia oocysts into a seedling raising pond for culturing.
The final hatching results of this comparative example were: oocysts that hatched for 4 days were difficult all reddened and white ulcerated, as shown in fig. 3, the larvae in the membrane died, the hatching rate was only 58%, and the survival rate was 64%.
Comparative example 2
The difference between this comparative example and example 5 is that: in the step (3), the buffer solution prepared in the example 1 is not used for soaking, and the Babylonia larvae are directly poured into a seedling raising pond. The final hatching results of this comparative example were: the hatching rate is 84 percent and the survival rate is 72 percent.
Comparative example 3
The difference between this comparative example and example 5 is that: in the step (1), the Dongfeng spiral oocysts are not punctured by using an oocone. The final hatching results of this comparative example were: the hatching rate is 68 percent and the survival rate is 85 percent.
Comparative example 4
The difference between this comparative example and example 5 is that: the buffer solution is not added with alpha-mangostin, and the final hatching result is as follows: the hatching rate is 90 percent and the survival rate is 78 percent.
Comparative example 5
The difference between this comparative example and example 5 is that: no florfenicol was added to the buffer, and the final incubation result was: the hatching rate is 88 percent and the survival rate is 75 percent.
Comparative example 6
The difference between this comparative example and example 5 is that: resveratrol is not added into the buffer solution, and the final incubation result is as follows: the hatching rate is 81 percent and the survival rate is 76 percent.
Comparative example 7
The difference between this comparative example and example 5 is that: the buffer solution components except for mangostin, resveratrol, florfenicol and dimethyl-beta-propiothiazole DMPT are not added, and the final hatching result is as follows: the hatching rate is 90 percent and the survival rate is 74 percent.
Test example 1
The testing method comprises the following steps: after 24 hours, randomly extracting 5 oocysts from the total oocysts for detecting fertilized egg quantity, observing under a microscope, recording the fertilized egg quantity of each oocyst, calculating to obtain an average value, and multiplying the average value by the total oocysts to obtain fertilized egg quantity. The oocyte development to the blastula stage is regarded as a successfully developed fertilized oosperm, and oocysts after the fertilization rate is measured are placed in a hatching frame again for further hatching and cultivation. After development at different stages, fertilized eggs begin to hatch out butterfly larvae. And (3) increasing the aeration quantity in the seedling raising pond, uniformly distributing butterfly larvae in the pond, then adopting a five-point sampling method to sample from 5 different positions of the seedling raising pond respectively by using a 100mL measuring cylinder, recording the number of larvae, and calculating the hatching rate.
The survival rate is the number of larvae 24 hours after the membrane rupture of the disc larvae on the inner surface of the membrane, compared with the number of larvae just at the time of membrane rupture, and the calculated number of larvae is also a five-point sampling method.
Fertilized egg quantity = fertilized egg quantity per oocyst x oocyst total.
Overall hatchability calculation formula: hatchability = number of larvae in late face plate/fertilized egg amount x 100%.
The overall survival rate was calculated as: survival = 24 hours later larvae/initial larvae hatched x 100%.
Final test data:
as is apparent from the above examples 5, 6, 1, 2 and 3, when the oocysts were punctured without using an egg-breaking cone or soaked without using the buffer prepared in example 1, the hatching rate and the survival rate of the hatching-difficult eastern-wind snail oocysts were greatly different from those of examples 5 and 6.
As can be seen from examples 5, 2, 4, 5 and 6, the survival rates of the eastern spirooocysts in comparative examples 4, 5 and 6 were lower when alpha-mangostin, florfenicol and resveratrol were absent, respectively, from the buffers. When the alpha-mangostin, the florfenicol and the resveratrol are simultaneously present in the buffer solution system, the survival rate of the hatching difficult eastern wind snail oocysts can reach 93.5 percent.
As is clear from comparative example 7, when the buffer system lacks magnesium glycinate, potassium chloride, calcium citrate, vitamin C, DL-tocopheryl acetate, proline, zinc amino acid complex, sodium selenate, ferrous fumarate, glucose, etc., the buffer is difficult to function to increase survival rate of the hatching-difficult Babylonia.
The foregoing examples are illustrative only and serve to explain some features of the method of the invention. The claims that follow are intended to claim the broadest possible scope as conceivable and the embodiments presented herein are demonstrated for the applicant's true test results. It is, therefore, not the intention of the applicant that the appended claims be limited by the choice of examples illustrating the features of the invention. Some numerical ranges used in the claims also include sub-ranges within which variations in these ranges should also be construed as being covered by the appended claims where possible.
Claims (9)
1. The application of a buffer solution for improving the hatching rate and the survival rate of the eastern snail oocysts in improving the hatching rate and the survival rate of the eastern snail oocysts is characterized in that the buffer solution comprises 0.1-0.3 g/L alpha-mangostin, 0.1-0.3 g/L resveratrol, 0.02-0.03 g/L florfenicol, 0.2-0.9 g/L dimethyl-beta-thiotepa DMPT, 1.0-5.0 g/L magnesium glycinate, 0.5-1.0 g/L potassium chloride, 1.0-5.0 g/L calcium citrate, 0.03-0.06 g/L vitamin C, 0.02-0.06 g/L alpha-tocopheryl acetate, 0.02-0.04 g/L proline, 0.01-0.03 g/L zinc amino acid complex, 0.01-0.03 g/L sodium selenate, 0.01-0.03 g/L ferrous fumarate and 0.01-0.03 g/L glucose.
2. The use according to claim 1, wherein the buffer comprises 0.2-0.3 g/L α -mangostin, 0.2-0.3 g/L resveratrol, 0.02-0.03 g/L florfenicol, 0.5-0.8 g/L dimethyl- β -propionic acid thiotepa DMPT, 3.0-4.0 g/L magnesium glycinate, 0.7-8.0 g/L potassium chloride, 4.0-5.0 g/L calcium citrate, 0.04-0.05 g/L vitamin C, 0.05-0.06 g/L DL-tocopheryl acetate, 0.03-0.04 g/L proline, 0.02-0.03 g/L zinc amino acid complex, 0.02-0.03 g/L sodium selenate, 0.02-0.03 g/L ferrous fumarate, 6.0-8.0 g/L glucose, 1L sterile seawater.
3. The use according to claim 1, wherein the buffer is prepared by the following method:
mixing sterilized seawater, alpha-mangostin, resveratrol, florfenicol, dimethyl-beta-thiotepa DMPT, magnesium glycinate, potassium chloride, calcium citrate, vitamin C, DL-tocopheryl acetate, proline, an amino acid zinc complex, sodium selenate, ferrous fumarate and glucose, and uniformly stirring to obtain the buffer solution for improving the hatching rate and the survival rate of the eastern-conch oocysts.
4. A method for hatching Babylonia oocysts, comprising the steps of:
s1, uniformly spreading eastern conch oocysts at the bottom of an incubation frame, and puncturing the eastern conch oocysts until all larvae can break membranes by themselves; or the eastern conch oocysts are evenly spread at the bottom of the hatching frame, and the hatching frame is placed under the air pressure, so that the eastern conch oocyst membranes are crushed under the air pressure;
s2, stopping oxygen for 5-20 minutes after 1-5 minutes of inflation, fishing up floating Babylonia larvae, and discarding the Babylonia larvae at the bottom layer;
s3, cleaning the floating Babylonia larvae in the step S2, mixing and soaking the Babylonia larvae, the sterilized seawater and the buffer solution for improving the hatching rate of the oocysts of the Babylonia for 1-3 hours, and then pouring the mixed solution into a seedling raising pond for raising seedlings;
the buffer solution comprises 0.1-0.3 g/L alpha-mangostin, 0.1-0.3 g/L resveratrol, 0.02-0.03 g/L florfenicol, 0.2-0.9 g/L dimethyl-beta-propionic acid thiopavilion DMPT, 1.0-5.0 g/L magnesium glycinate, 0.5-1.0 g/L potassium chloride, 1.0-5.0 g/L calcium citrate, 0.03-0.06 g/L vitamin C, 0.02-0.06 g/L LDL-tocopheryl acetate, 0.02-0.04 g/L proline, 0.01-0.03 g/L amino acid zinc complex, 0.01-0.03 g/L sodium selenate, 0.01-0.03 g/L ferrous fumarate, 5.0-10.0 g/L glucose and 1L sterile seawater.
5. The method of hatching the eastern conch oocysts according to claim 4, wherein the eastern conch oocysts are mature eastern conch oocysts, larvae inside the oocysts have developed into pand larvae inside the membrane, the oocysts are light yellow to earthy yellow in appearance and finally dark brown in color, the edges are blackened, and the oocyst endosperm liquid is in a saturated state from a viscous state to a water-like state.
6. The method according to claim 4, wherein the concentration of the buffer for increasing the hatching rate of the eastern conch oocysts in the step S3 is 3-10 ppm.
7. The method of hatching an oocyst of Babylonia according to claim 4, wherein said Babylonia is selected from one or more of Babylonia quadrangularis, babylonia taiwan or Babylonia.
8. The method according to claim 4, wherein in the step S3, the density of the Babylonia larvae in the breeding pond is 60-120/L.
9. The method of hatching Babylonia oocysts according to claim 4, wherein in step S1, the air pressure is in the range of 200 to 400 kPa.
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