CN108935442A - 一种绵羊精液低温保存技术 - Google Patents
一种绵羊精液低温保存技术 Download PDFInfo
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- 241001494479 Pecora Species 0.000 title claims abstract description 36
- 238000005516 engineering process Methods 0.000 title claims abstract description 11
- 210000000582 semen Anatomy 0.000 title abstract description 15
- 239000007788 liquid Substances 0.000 claims abstract description 21
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000005138 cryopreservation Methods 0.000 claims abstract description 12
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims abstract description 6
- 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 6
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 claims abstract description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 6
- 229930006000 Sucrose Natural products 0.000 claims abstract description 6
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000008103 glucose Substances 0.000 claims abstract description 6
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 claims abstract description 6
- 229940083466 soybean lecithin Drugs 0.000 claims abstract description 6
- 239000005720 sucrose Substances 0.000 claims abstract description 6
- 235000020183 skimmed milk Nutrition 0.000 claims abstract description 4
- 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 3
- 230000003115 biocidal effect Effects 0.000 claims description 3
- 239000002131 composite material Substances 0.000 claims description 3
- 230000004913 activation Effects 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000002577 cryoprotective agent Substances 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 12
- 238000004321 preservation Methods 0.000 abstract description 9
- 238000009395 breeding Methods 0.000 abstract description 3
- 230000001488 breeding effect Effects 0.000 abstract description 3
- 239000003242 anti bacterial agent Substances 0.000 abstract 1
- 229940088710 antibiotic agent Drugs 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 abstract 1
- 244000144972 livestock Species 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 239000000843 powder Substances 0.000 abstract 1
- 239000003761 preservation solution Substances 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 10
- 230000009027 insemination Effects 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- 230000000694 effects Effects 0.000 description 3
- 230000008021 deposition Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 210000001215 vagina Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000628997 Flos Species 0.000 description 1
- 208000022555 Genital disease Diseases 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000009402 cross-breeding Methods 0.000 description 1
- 230000002338 cryopreservative effect Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000003507 refrigerant Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000019100 sperm motility Effects 0.000 description 1
- 239000002699 waste material Substances 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
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
-
- 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
- A01N1/0215—Disinfecting agents, e.g. antimicrobials for preserving living parts
-
- 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
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- Health & Medical Sciences (AREA)
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Abstract
本发明涉及一种绵羊精液低温保存技术，具体涉及绵羊精液低温保存液配方和绵羊精液低温保存操作方法，属家畜繁殖技术领域。保存液配方的化学组成：脱脂奶粉，棉籽糖，葡萄糖，蔗糖，大豆卵磷脂，甘油和复合抗生素。一种绵羊精液低温保存技术的具体操作方法：用本配方液对合格绵羊精液稀释15倍，放于外包裹20层纱布的离心管中，于0‑4℃保存1小时拆掉纱布，继续于0‑4℃保存，有效期为6.5天。该发明与其他绵羊精液低温保存技术相比较，具有精液保存时间长、受胎率高等优点；与绵羊精液冷冻保存技术相比较，具有储存和运输条件要求低等优点。
Description
技术领域
本发明涉及一种绵羊精液低温保存技术，具体涉及绵羊精液低温保存液配方和绵羊精液低温保存操作方法。
背景技术
在畜牧生产实践中，羊人工授精技术可在短期内最大程度地发挥优秀种公羊的遗传潜力、加速杂交改良、加快育种进程、防止疾病传播，特别是在生殖道疾病的传播等方面具有重要意义，而精液保存效果优劣、储存运输要求条件难易会在很大程度上影响羊人工授精技术的推广应用。目前，保存绵羊精液常见的方法是冷冻保存，可实现绵羊精液的长期保存，但使用绵羊冷冻精液进行人工授精的效果并不理想，问题突出表现为：①阴道输精的受胎率低（30%-50%），这与绵羊精子对冷冻打击异常敏感、冻融过程很容易对精子形态和运动能力造成损伤等因素有关；②储存和运输要求条件高，绵羊精液冷冻保存的必要条件是精液始终浸泡在-196℃的液氮中，所需特殊保存容器（液氮罐）的价格高，并且要求及时添加液氮（偏僻地区不易获得），另外在崎岖山路运输时的强烈颠簸并不能很好地保障冻精品质，也不便于经常转运分发到各个养殖场或牧区配种站。
绵羊精液低温保存技术，是指将合格的新鲜绵羊精液用本发明的保存液稀释后在0-4℃条件下保存一段时间，在进行绵羊人工授精时可将处于保存有效期内的绵羊精液进行复温处理，使绵羊精子依然具有正常的受精能力。原理：绵羊精液于0-4℃条件下保存，精子几乎处于休眠状态，消耗营养量低，代谢废物累积量少，可有效延长精子存活时间。
该发明采用将甘油和大豆卵磷脂配制成B液的技术工艺提高了精子对冷打击的耐受能力，利用M199作为精液复温后的一种激活剂解决了所保存精子活力受到酸抑制的问题，将棉籽糖、葡萄糖、蔗糖进行组合应用则能够更好地满足精子存活对能量的需求，因此具有精液低温保存时间长（6.5天）、阴道输精的受胎率高（70%以上）等优点。与绵羊精液冷冻保存技术相比较，本技术不需要价格昂贵的液氮罐及特殊冷媒（液氮），储存和运输要求条件低。
发明内容
提供一种绵羊精液低温保存液配方，100ml保存液含：脱脂奶粉10g、棉籽糖0.25g、葡萄糖0.75g、蔗糖0.75g、大豆卵磷脂0.375g、甘油0.75ml和复合抗生素2ml，以及一种绵羊精液低温保存的操作方法。
具体实施方式
以下实施例用于说明本发明，包括以下步骤：
a、将脱脂奶粉10g、棉籽糖0.25g、葡萄糖0.75g、蔗糖0.75g和复合抗生素2ml溶解于48ml经过高压灭菌的蒸馏水中，制成A液。
b、将0.375g大豆卵磷脂溶解于0.75ml的甘油中，配制成B液。
c、将A液加入B液中，然后用49.25ml经高压灭菌的蒸馏水冲刷A液容器并移至到B液中，并充分搅拌混匀，即制成绵羊精液低温保存液。
d、原精活力达到70%以上的绵羊鲜精，用本保存液进行15倍稀释，放于外包裹20层纱布的离心管中，于0-4℃条件下保存1小时拆掉纱布，将装有绵羊精液的离心管于0-4℃继续保存，有效保存期为6.5天。
e、绵羊进行人工输精时，将经低温保存的绵羊精液于38℃水浴锅中复温30s。
f、向复温后的绵羊精液中加入与同体积的等温M199并混匀，即可用于绵羊人工输精。
Claims (2)
1.一种绵羊精液低温保存技术，其保存液每100ml包括如下组分：脱脂奶粉10g、棉籽糖0.25g、葡萄糖0.75g、蔗糖0.75g、大豆卵磷脂0.375g、甘油0.75ml和复合抗生素2ml；特征在于其技术方案：甘油和大豆卵磷脂作为低温保护剂组合并配制成B液的技术工艺，M199用作精液复温后的一种精子激活剂，棉籽糖、葡萄糖、蔗糖作为能量物质的组合应用。
2.权利要求1中所述的绵羊精液低温保存液配方及其技术应用。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109497044A (zh) * | 2018-12-28 | 2019-03-22 | 深圳市昱杰生物科技有限公司 | 一种哺乳动物早期胚胎冻存液及冻存方法 |
| CN113016779A (zh) * | 2021-02-07 | 2021-06-25 | 内蒙古大学 | 一种奶绵羊精液冷冻保存方法及其稀释液 |
| CN115624024A (zh) * | 2022-08-22 | 2023-01-20 | 甘肃润牧生物工程有限责任公司 | 党参多糖作为添加剂在绵羊精液低温稀释液中的应用 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109497044A (zh) * | 2018-12-28 | 2019-03-22 | 深圳市昱杰生物科技有限公司 | 一种哺乳动物早期胚胎冻存液及冻存方法 |
| CN113016779A (zh) * | 2021-02-07 | 2021-06-25 | 内蒙古大学 | 一种奶绵羊精液冷冻保存方法及其稀释液 |
| CN115624024A (zh) * | 2022-08-22 | 2023-01-20 | 甘肃润牧生物工程有限责任公司 | 党参多糖作为添加剂在绵羊精液低温稀释液中的应用 |
| CN115624024B (zh) * | 2022-08-22 | 2023-08-22 | 甘肃润牧生物工程有限责任公司 | 党参多糖作为添加剂在绵羊精液低温稀释液中的应用 |
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