CN109329140A - A kind of green Medaka incubating oosperm liquid of simplification - Google Patents
A kind of green Medaka incubating oosperm liquid of simplification Download PDFInfo
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- CN109329140A CN109329140A CN201811540469.9A CN201811540469A CN109329140A CN 109329140 A CN109329140 A CN 109329140A CN 201811540469 A CN201811540469 A CN 201811540469A CN 109329140 A CN109329140 A CN 109329140A
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- nahco
- green medaka
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- 241000276569 Oryzias latipes Species 0.000 title claims abstract description 62
- 239000007788 liquid Substances 0.000 title claims abstract description 44
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 22
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 16
- 235000013601 eggs Nutrition 0.000 abstract description 41
- 239000012530 fluid Substances 0.000 abstract description 38
- 230000012447 hatching Effects 0.000 abstract description 33
- 238000011534 incubation Methods 0.000 abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 13
- 238000004043 dyeing Methods 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 6
- 230000008569 process Effects 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 34
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 239000008399 tap water Substances 0.000 description 8
- 235000020679 tap water Nutrition 0.000 description 8
- 239000001110 calcium chloride Substances 0.000 description 7
- 229910001628 calcium chloride Inorganic materials 0.000 description 7
- 201000010099 disease Diseases 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 244000137852 Petrea volubilis Species 0.000 description 6
- 239000001103 potassium chloride Substances 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 5
- 238000005273 aeration Methods 0.000 description 5
- 241000251468 Actinopterygii Species 0.000 description 4
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 3
- 229910052564 epsomite Inorganic materials 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 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 description 2
- 210000004392 genitalia Anatomy 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 229960000907 methylthioninium chloride Drugs 0.000 description 2
- 238000005096 rolling process Methods 0.000 description 2
- 239000004576 sand Substances 0.000 description 2
- 230000007480 spreading Effects 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 210000003484 anatomy Anatomy 0.000 description 1
- 239000003855 balanced salt solution Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 231100000613 environmental toxicology Toxicity 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M potassium chloride Inorganic materials [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- 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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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Environmental Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Inorganic Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A kind of green Medaka incubating oosperm liquid of simplification, it is related to a kind of incubation fluid.The present invention in order to solve the problems, such as existing incubation fluid be easy dyeing, complicated component, exist to user it is certain dangerous.The green Medaka incubating oosperm liquid that the present invention simplifies is the NaHCO of basicity 30mM~50mM3The NaHCO of solution or pH value 9~123Solution.Incubation fluid of the invention can successfully hatch green Medaka seedling, and the morbidity of green Medaka fertilized eggs water mo(u)ld can also be treated in hatching process, solve the problems, such as that existing incubation fluid is easy dyeing, complicated component, has certain potential hazard to the body of operator.
Description
Technical field
The present invention relates to a kind of incubation fluid, in particular to a kind of incubation fluid of green Medaka fertilized eggs.
Background technique
Japanese blueness Medaka (Oryzias latipes) is a kind of fresh water QingMedaka section fish, be distributed widely in Japan, Korea, in
The countries and regions such as state east, the southeast and Southeast Asia.Since green Medaka has, individual is small, the easy differentiation of gender, the generation cycle is short, produces
The features such as ovum rate is high, raising breeding is simple, genomic information understands, thus usually using it as one kind be used for fish anatomy,
The model organism that embryology, physiology, gene function are verified.In addition, medaka is that International Standards Organization (ISO) recommends toxicity examination
The standard tested is widely used in field of environmental toxicology with one of fish.Green Medaka fertilized eggs are widely used in life science
Every field, enough fertilized eggs can be obtained, cultivating into seedling and adult fish is basis and the premise for carrying out correlation test.
The green Medaka incubating oosperm liquid of standard generally includes following chemical component: 1g/L NaCl, 0.03g/L KCl,
0.04g/L CaCl2·2H2O and 0.16g/L MgSO47H2O, in order to effectively prevent the growth of water mo(u)ld, usually in standard
0.01% methylene blue solution (10ml/L) is added in liquid.But since methylene blue is easy to dye, often to experiment
Operator brings unnecessary trouble, as the short time is not easy to wash off after clothing dyeing or finger dyeing.In lot of documents usually
The incubation fluid referred to further include: Yamamoto ' s Ringer ' s solution, ingredient include 0.75%NaCl, 0.02%KCl,
0.02%CaCl2, 0.002%NaHCO3, adjust pH 7.3 (Yamamoto T.1939.Proc.Imp.Acad.Tokyo, 15,
269-271)；Solution similar with Yamamoto ' s Ringer ' s incubation fluid further includes Iwamatsu ' s balanced salt
Solution, abbreviation Iwamatsu ' s BSS (Iwamatsu T.1983.J.Exp.Zool.228,83-89), ingredient includes
6.5g/L NaCl, 0.4g/L KCl, 0.2g/L CaCl2·2H2O, 0.2g/L MgSO4·7H2O uses 5%NaHCO3It adjusts
PH to 7.4.It can be seen that no matter being needed a variety of in use using standard hatching liquid or other two kinds of incubation fluids
Chemical reagent NaCl, KCl, CaCl2And MgSO4, comparison of ingredients is complicated, wherein CaCl2It is also easy to make moist, causes dosage inaccurate
Really, MgSO4The corrosivity having, to user, there are certain potential hazards.
Summary of the invention
The easy dyeing of existing incubation fluid, complicated component, to user, there are certain potential hazards in order to solve by the present invention
The problem of, and a kind of green Medaka incubating oosperm liquid of the simplification provided.
The green Medaka incubating oosperm liquid that the present invention simplifies is the NaHCO of basicity 30mM~50mM3Solution.
The green Medaka incubating oosperm liquid that the present invention simplifies is the NaHCO of pH value 9~123Solution.
Solution total alkalinity formula=HCO3 -+2(CO3 2-)+OH-+H+.When pH is 6.35~10.33, HCO in solution3 -It is
Basicity is primarily present form；When pH is higher than 10.33, CO in solution3 2-It is main component (the Thomas S.& of basicity
Poupin J.1985.J.Comp.Physiol.B.156,29-34).Incubation fluid of the present invention is NaHCO3Solution improves hatching
The total alkalinity of liquid, buffer capacity is good, and living environment is more stable mild for green Medaka fertilized eggs, is more advantageous to fertilized eggs
Growth.Hatch green Medaka fertilized eggs using incubation fluid of the invention, hatching rate is 50%~55%.Incubation fluid of the invention can succeed
Hatch green Medaka, the morbidity of the water mo(u)ld of green Medaka fertilized eggs can also be treated in hatching process, the disease incidence of water mo(u)ld is
24%~30%.Green Medaka incubating oosperm liquid of the invention is NaHCO3, formula is simple, can guarantee the hatching of green Medaka, also solves
Existing incubation fluid is easy dyeing, complicated component, there is a problem of danger centainly to user.
Specific embodiment
Specific embodiment 1: the green Medaka incubating oosperm liquid that present embodiment simplifies is basicity 30mM~50mM's
NaHCO3Solution.
Hatch green Medaka fertilized eggs using the incubation fluid of present embodiment, average percentage hatch rate is 50%~55%, is being hatched
The disease incidence of water mo(u)ld is 24%~30% in journey, and incubation fluid of the invention can successfully hatch green Medaka seedling, hatch
It also can control the water mo(u)ld of green Medaka fertilized eggs in journey.
Specific embodiment 2: the present embodiment is different from the first embodiment in that: the blueness that present embodiment simplifies
Medaka incubating oosperm liquid is the NaHCO of basicity 35mM~45mM3Solution.Other are same as the specific embodiment one.
Specific embodiment 3: the present embodiment is different from the first embodiment in that: the blueness that present embodiment simplifies
The NaHCO of Medaka incubating oosperm liquid alkaline degree 40mM3Solution.Other are same as the specific embodiment one.
Specific embodiment 4: the green Medaka incubating oosperm liquid that present embodiment simplifies is the NaHCO of pH value 9~123It is molten
Liquid.
Hatch green Medaka fertilized eggs using the incubation fluid of present embodiment, average percentage hatch rate is 50%~55%, is being hatched
The disease incidence of water mo(u)ld is 24%~30% in journey, and incubation fluid of the invention can successfully hatch green Medaka, in hatching process
It also can control the water mo(u)ld of green Medaka fertilized eggs.
Specific embodiment 5: present embodiment is unlike specific embodiment four: the blueness that present embodiment simplifies
Medaka incubating oosperm liquid is the NaHCO of pH value 10~113Solution.Other are identical as specific embodiment four.
Specific embodiment 6: present embodiment is unlike specific embodiment four: the blueness that present embodiment simplifies
Medaka incubating oosperm liquid is the NaHCO of pH value 10.33Solution.Other are identical as specific embodiment four.
Embodiment 1
(i.e. basicity is 30,40 and 50mM NaHCO to the present invention 3 different basicity blueness Medaka incubating oosperm liquid3Solution) and mark
Quasi- incubation fluid (1g/L NaCl, 0.03g/L KCl, 0.04g/L CaCl2·2H2O,0.16g/L MgSO4·7H2O), it is aerated certainly
Experiment is compared in water, and the specific method is as follows:
Step 1: configuration basicity is 30,40 and 50mM NaHCO3Solution and standard hatching liquid；
Step 2: spreading the sand paper of 2000 mesh in culture dish bottom, and frosted is face-up, and aeration tap water is added, submerges sand
Paper, and liquid level is away from sand paper upper surface distance≤1mm, wherein the diameter of culture dish be 8.7cm, depth 1.2cm, solution deep is
1.0cm or so)；
Step 3: after green Medaka is laid eggs 2~3 hours, thyrsiform fertilized eggs are shelled from female genital orifice using hairbrush
From and be collected together, collect female fertilized eggs grain；
Step 4: the female fertilized eggs grain collected in step 3 is transferred to the wet sand of step 2 with suction pipe respectively
Above paper, female fertilized eggs grain 40 times or so of rolling are rubbed, it is made to be separated into single fertilized eggs；
Step 5: the fertilized eggs of step 4 are taken to be transferred in 5 groups of different hatching test fluids respectively with suction pipe, first group incubates
Changing test fluid is 30mM NaHCO3Solution, second group of hatching test fluid are 40mM NaHCO3Solution, third group hatching test fluid are
50mM NaHCO3Solution, the 4th group of hatching test fluid are standard hatching liquid, and the 5th group of hatching test fluid is to be aerated tap water, five
Group sample, which is placed in 26 DEG C of incubator, to be cultivated, and every 6h observes fertilized eggs, chooses the fertilized eggs with water mold,
Hatching test fluid is replaced daily.
By experimental observation, basicity 30mM NaHCO is used3The green Medaka fertilized eggs that solution is hatched, started at 12.3 days
Hatching；Basicity 40mM NaHCO3The green Medaka fertilized eggs that solution is hatched, started to hatch at 13 days, basicity 50mM NaHCO3
The green Medaka fertilized eggs that solution is hatched, started to hatch at 13.5 days.The green Medaka fertilized eggs hatched using standard hatching liquid,
Started to hatch at 12.5 days.The green Medaka fertilized eggs hatched using aeration tap water, started to hatch at 12.5 days.
The experimental result of the present embodiment is as shown in table 1.
Table 1
| Hatch test fluid | Hatching rate (%) | Saprolegniasis disease incidence (%) |
| 30mM NaHCO3 | 50 | 28 |
| 40mM NaHCO3 | 52.5 | 26 |
| 50mM NaHCO3 | 50 | 25 |
| Standard hatching liquid | 55 | 27 |
| It is aerated tap water | 27 | 55 |
It can be seen that the hatchability of incubation fluid of the present invention and the hatchability ten of standard hatching liquid from the data in table 1
Tap is close, has reached 50%~52.5%, can successfully hatch green Medaka using incubation fluid of the invention, and use the present invention
The disease incidence of the green Medaka fertilized eggs water mo(u)ld of incubation fluid is 25%~28%, well below what is hatched using aeration tap water
Green Medaka fertilized eggs, incubation fluid of the invention have certain therapeutic effect to the saprolegniasis of green Medaka fertilized eggs；By to data in table 1
Observation, uses 30~50mM of basicity NaHCO3Solution may make the hatching rate of green Medaka fertilized eggs reach 50% or more, water mold hair
Sick rate is lower than 30%.
Embodiment 2
Green Medaka incubating oosperm liquid (the i.e. NaHCO that pH is 9.3,10.3 and 11.3 of the present invention 3 different pH value3Solution)
With standard hatching liquid (1g/L NaCl, 0.03g/L KCl, 0.04g/L CaCl2·2H2O, 0.16g/L MgSO4·7H2O), expose
Experiment is compared in gas tap water, and the specific method is as follows:
Step 1: the NaHCO that configuration pH is 9.3,10.3 and 11.33Solution (10mM) and standard hatching liquid；
Step 2: spreading the sand paper of 2000 mesh in culture dish bottom, and frosted is face-up, and aeration tap water is added, submerges sand
Paper, and liquid level is away from sand paper upper surface distance≤1mm, wherein the diameter of culture dish be 8.7cm, depth 1.2cm, solution deep is
1.0cm or so)；
Step 3: after green Medaka is laid eggs 2~3 hours, thyrsiform fertilized eggs are shelled from female genital orifice using hairbrush
From and be collected together, collect female fertilized eggs grain；
Step 4: the female fertilized eggs grain collected in step 3 is transferred to the wet sand of step 2 with suction pipe respectively
Above paper, rolling fertilized eggs grain 40 times or so is rubbed, it is made to be separated into single fertilized eggs；
Step 5: the fertilized eggs of step 4 are taken to be transferred in 5 groups of different hatching test fluids respectively with suction pipe, first group incubates
Change test fluid is that pH is 9.3NaHCO3Solution, second group of hatching test fluid are that pH is 10.3NaHCO3Solution, the hatching of third group are surveyed
Test solution is that pH is 11.3NaHCO3Solution, the 4th group of hatching test fluid are standard hatching liquid, and the 5th group of hatching test fluid is aeration
Tap water, five groups of samples are placed in 26 DEG C of incubator and are cultivated, and every 6h observes fertilized eggs, chooses with water mold
Fertilized eggs, incubation fluid is replaced daily.
The experimental result of the present embodiment is as shown in table 2.
Table 2
It can be seen that the NaHCO that pH is 9.3,10.3 and 11.3 from the data in table 23Solution, saprolegniasis disease incidence are close
Standard hatching liquid, the NaHCO of pH 9.3,10.3 and 11.33Solution hatching rate 50%~55% and saprolegniasis disease incidence 24%
~30%；Incubation fluid of the invention can hatch green Medaka fertilized eggs, and have certain treatment to make the saprolegniasis of green Medaka fertilized eggs
With.
It is compared by Tables 1 and 2, it was found that as basicity and pH value increase, hatching rate is improved.But work as basicity liter
High to be increased to 11.3 to 50mM and pH value, hatching rate decline, most suitable basicity is 40mM and pH value is 10.3.Measurement 30,40,50mM
The NaHCO of basicity3After the pH of solution, it is found that the pH of these gradient alkalinity solutions is respectively 8.33,8.34 and 8.39, it is several between pH
There is no difference.
Claims (6)
1. a kind of green Medaka incubating oosperm liquid of simplification, it is characterised in that simplify green Medaka incubating oosperm liquid be basicity 30mM~
The NaHCO of 50mM3Solution.
2. blueness Medaka incubating oosperm liquid according to claim 1, it is characterised in that the green Medaka incubating oosperm liquid simplified is
The NaHCO of basicity 35mM~45mM3Solution.
3. blueness Medaka incubating oosperm liquid according to claim 1, it is characterised in that the green Medaka incubating oosperm liquid simplified is
The NaHCO of basicity 40mM3Solution.
4. a kind of green Medaka incubating oosperm liquid of simplification, it is characterised in that the green Medaka incubating oosperm liquid simplified is pH value 9~12
NaHCO3Solution.
5. blueness Medaka incubating oosperm liquid according to claim 4, it is characterised in that the green Medaka incubating oosperm liquid simplified is
The NaHCO of pH value 10~113Solution.
6. blueness Medaka incubating oosperm liquid according to claim 4, it is characterised in that the green Medaka incubating oosperm liquid simplified is
The NaHCO of pH value 10.33Solution.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111034656A (en) * | 2019-11-24 | 2020-04-21 | 南通科技职业学院 | Preparation method for preparing hatching enzyme solution based on medaka hatching fluid |
| CN111248115A (en) * | 2020-01-10 | 2020-06-09 | 山东省海洋生物研究院 | Indoor culture method for obtaining high-quality fertilized eggs of seawater medaka |
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| CN105007990A (en) * | 2012-12-21 | 2015-10-28 | 阿克生物科技公司 | A cosmetic composition from fish hatching fluid, methods for its production and uses thereof for improving the cosmetic appearance of skin |
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| Title |
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| YAO Z L,LAI Q F,ZHOU K,ET AL: "《Developmental biology of medaka fish (Oryzias latipes) exposed to alkalinity stress》", 《JOURNAL OF APPLIED ICHTHYOLOGY》 * |
| 张静 等: "《青鳉（Oryzias latipes）性腺分化与发育的组织学观察》", 《动物学研究》 * |
| 徐敏: "《盐碱度对3种鱼精子活力、受精率及孵化率的影响》", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111034656A (en) * | 2019-11-24 | 2020-04-21 | 南通科技职业学院 | Preparation method for preparing hatching enzyme solution based on medaka hatching fluid |
| CN111248115A (en) * | 2020-01-10 | 2020-06-09 | 山东省海洋生物研究院 | Indoor culture method for obtaining high-quality fertilized eggs of seawater medaka |
| CN111248115B (en) * | 2020-01-10 | 2021-10-26 | 山东省海洋生物研究院 | Indoor culture method for obtaining high-quality fertilized eggs of seawater medaka |
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