CN105557683A - 一种泥鳅精子的保存方法及相应的泥鳅精子保存液 - Google Patents
一种泥鳅精子的保存方法及相应的泥鳅精子保存液 Download PDFInfo
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- CN105557683A CN105557683A CN201610026066.7A CN201610026066A CN105557683A CN 105557683 A CN105557683 A CN 105557683A CN 201610026066 A CN201610026066 A CN 201610026066A CN 105557683 A CN105557683 A CN 105557683A
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- 241000252185 Cobitidae Species 0.000 title claims abstract description 63
- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000003879 sperm preservation Methods 0.000 title abstract 6
- 239000003761 preservation solution Substances 0.000 title abstract 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000012153 distilled water Substances 0.000 claims abstract description 7
- 230000009027 insemination Effects 0.000 claims abstract description 7
- 239000012530 fluid Substances 0.000 claims description 14
- 239000003755 preservative agent Substances 0.000 claims description 14
- 230000002335 preservative effect Effects 0.000 claims description 14
- 102000002322 Egg Proteins Human genes 0.000 claims description 6
- 108010000912 Egg Proteins Proteins 0.000 claims description 6
- 210000004681 ovum Anatomy 0.000 claims description 6
- 230000009303 sperm storage Effects 0.000 claims description 3
- 238000005273 aeration Methods 0.000 claims description 2
- 230000012447 hatching Effects 0.000 claims description 2
- 238000004321 preservation Methods 0.000 abstract description 5
- 230000004083 survival effect Effects 0.000 abstract description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 abstract 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 abstract 1
- 239000007836 KH2PO4 Substances 0.000 abstract 1
- -1 Na2HPO4.7H2O Substances 0.000 abstract 1
- 239000001110 calcium chloride Substances 0.000 abstract 1
- 235000011148 calcium chloride Nutrition 0.000 abstract 1
- 229910001628 calcium chloride Inorganic materials 0.000 abstract 1
- DHRRIBDTHFBPNG-UHFFFAOYSA-L magnesium dichloride hexahydrate Chemical compound O.O.O.O.O.O.[Mg+2].[Cl-].[Cl-] DHRRIBDTHFBPNG-UHFFFAOYSA-L 0.000 abstract 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 abstract 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 abstract 1
- 235000019796 monopotassium phosphate Nutrition 0.000 abstract 1
- 239000001103 potassium chloride Substances 0.000 abstract 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 abstract 1
- 235000017557 sodium bicarbonate Nutrition 0.000 abstract 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 abstract 1
- 239000011780 sodium chloride Substances 0.000 abstract 1
- 230000004720 fertilization Effects 0.000 description 4
- 210000000582 semen Anatomy 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- Marine Sciences & Fisheries (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种泥鳅精子的保存方法及相应的泥鳅精子保存液，该保存液是将KCl、NaCl、Na2HPO4·7H2O、KH2PO4、CaCl2、MgSO4·7H2O、MgCl2·6H2O溶于蒸馏水中，使用前加入NaHCO3，加蒸馏水定容而成。本发明所述泥鳅精子的保存方法是将泥鳅精子与泥鳅精子保存液混合于培养皿中，避光保存；所述保存方法可以完好的大量保存泥鳅精子，提高了精子的存活率，保存方法简单，为泥鳅人工授精提供强力支持，适合在本领域推广应用。
Description
技术领域
本发明涉及生物领域，具体涉及泥鳅精子的保存方法及相应的泥鳅精子保存液。
技术背景
泥鳅繁殖具有受精率低的特点，所以人工泥鳅繁殖需要大量的泥鳅精子，但泥鳅精子又存在不易保存的问题亟待解决。现有的泥鳅精子保存时多数都是在-5℃或更低的温度下保存，给实际人工繁殖带来不便。
发明内容
本发明的目的在于克服上述缺点，提供一种在4℃即可妥善保存泥鳅精子的保存方法。
本发明另一目的在于提供上述保存泥鳅精子的保存液。
本发明还提供了利用泥鳅精子保存液保存好的泥鳅精子与泥鳅卵子人工授精的方法。
本发明上述目的通过以下技术方案予以实现：
一种泥鳅精子保存液，是将0.35gKCl、6.2gNaCl、0.7gNa2HPO4·7H2O、0.65gKH2PO4、1.2gCaCl2、1.25gMgSO4·7H2O、0.1gMgCl2·6H2O充分溶于900ml蒸馏水中，使用前加入0.4gNaHCO3，加蒸馏水定容至1L而成。
一种泥鳅精子保存方法，是将泥鳅精子与4℃预冷的泥鳅精子保存液按照1:30~1:50的体积比混合均匀于容器中，4℃避光保存。
将本发明4℃避光保存的泥鳅精液与泥鳅卵子按照体积比为1:10的比例在干净容器中混匀，混匀后再转入经充分曝气的自来水中清洗2~3次，进行循环水孵化，从而实现泥鳅人工授精。
与现有技术相比，本发明具有如下有益效果：
（1）本发明所述泥鳅精子的保存方法可以完好的大量保存泥鳅精子，提高了精子的存活率，保存方法简单，为泥鳅人工授精提供强力支持，适合在本领域推广应用；
（2）本发明保存温度为4℃，保存温度无需-5℃以下，避免反复解冻对精子带来的损害，为泥鳅人工繁殖提供很大的便利。
具体实施方式
下面结合实施例，对本发明做进一步阐述。
实施例1
泥鳅精子保存液是将0.35gKCl、6.2gNaCl、0.7gNa2HPO4·7H2O、0.65gKH2PO4、1.2gCaCl2、1.25gMgSO4·7H2O、0.1gMgCl2·6H2O充分溶于900ml蒸馏水中，使用前加入0.4gNaHCO3，加蒸馏水定容至1L而成。
将泥鳅精子与4℃预冷的泥鳅精子保存液按照1:30的体积比混合均匀于容器中，4℃避光保存2小时后，使用普通光学显微镜检查泥鳅精子的活性，具运动能力的泥鳅精子占比为88.26%以上。使用上述泥鳅精液与泥鳅卵子按照体积比为1:10的比例进行人工授精，受精率达到88.40%。
实施例2
泥鳅精子保存液是将0.35gKCl、6.2gNaCl、0.7gNa2HPO4·7H2O、0.65gKH2PO4、1.2gCaCl2、1.25gMgSO4·7H2O、0.1gMgCl2·6H2O充分溶于900ml蒸馏水中，使用前加入0.4gNaHCO3，加蒸馏水定容至1L而成。
将泥鳅精子与4℃预冷的泥鳅精子保存液按照1:40的体积比混合均匀于容器中，4℃避光保存2小时后，使用普通光学显微镜检查泥鳅精子的活性，具运动能力的泥鳅精子占比为90.05%以上。使用上述泥鳅精液与泥鳅卵子按照体积比为1:10的比例进行人工授精，受精率达到91.12%。
实施例3
泥鳅精子保存液是将0.35gKCl、6.2gNaCl、0.7gNa2HPO4·7H2O、0.65gKH2PO4、1.2gCaCl2、1.25gMgSO4·7H2O、0.1gMgCl2·6H2O充分溶于900ml蒸馏水中，使用前加入0.4gNaHCO3，加蒸馏水定容至1L而成。
将泥鳅精子与4℃预冷的泥鳅精子保存液按照1:50的体积比混合均匀于容器中，4℃避光保存2小时后，使用普通光学显微镜检查泥鳅精子的活性，具运动能力的泥鳅精子占比为89.36%以上。使用上述泥鳅精液与泥鳅卵子按照体积比为1:10的比例进行人工授精，受精率达到89.23%。
Claims (3)
1.一种泥鳅精子保存液，其特征在于所述泥鳅精子保存液是将0.35gKCl、6.2gNaCl、0.7gNa2HPO4·7H2O、0.65gKH2PO4、1.2gCaCl2、1.25gMgSO4·7H2O、0.1gMgCl2·6H2O充分溶于900ml蒸馏水中，使用前加入0.4gNaHCO3，加蒸馏水定容至1L而成。
2.一种泥鳅精子保存方法，其特征在于所述保存方法是将泥鳅精子与4℃预冷的权利要求1所述泥鳅精子保存液按照1:30~50的体积比混合均匀于容器中，4℃避光保存。
3.一种泥鳅人工授精方法，其特征在于将权利要求2所述保存的泥鳅精子，与泥鳅卵子按照1:10的体积比混匀后，转入经充分曝气的自来水中清洗2~3次，进行循环水孵化。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107683798A (zh) * | 2017-09-27 | 2018-02-13 | 清远职业技术学院 | 一种黄颡鱼人工育苗和饲养的方法 |
| CN108552161A (zh) * | 2018-05-22 | 2018-09-21 | 华中农业大学 | 一种泥鳅精子的超低温冷冻保存液及其冷冻复苏方法和应用 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107683798A (zh) * | 2017-09-27 | 2018-02-13 | 清远职业技术学院 | 一种黄颡鱼人工育苗和饲养的方法 |
| CN108552161A (zh) * | 2018-05-22 | 2018-09-21 | 华中农业大学 | 一种泥鳅精子的超低温冷冻保存液及其冷冻复苏方法和应用 |
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Application publication date: 20160511 |