CN111685102A - Sheep sperm solution diluent powder - Google Patents
Sheep sperm solution diluent powder Download PDFInfo
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- CN111685102A CN111685102A CN202010383699.XA CN202010383699A CN111685102A CN 111685102 A CN111685102 A CN 111685102A CN 202010383699 A CN202010383699 A CN 202010383699A CN 111685102 A CN111685102 A CN 111685102A
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- sheep semen
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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
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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
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to sheep semen diluent powder, which belongs to the field of animal reproduction and comprises the following components in formula: 0.1-0.3g of monoammonium glycyrrhizinate, 0.5-1.0g of glutamic acid, 0.5-1.0g of L-arginine and 0.5-1.0g of melatonin; 37-40g of glucose, 4-8g of citric acid, 1-2g of EDTA, 1-2g of sodium carbonate and 0.1-0.3g of oregano oil, and the diluent powder and PBS buffer solution are prepared into 1L of diluent, so that the time and the survival rate of the preserved sperms can be obviously improved, the integrity rate of acrosomes is improved, and the conception rate of diluted artificial insemination is improved.
Description
Technical Field
The invention belongs to the technical field of animal breeding, and particularly relates to sheep sperm solution diluted powder.
Background
With the continuous improvement of the income level of residents in China, the consumption concept and the living style are continuously changed, and the consumption of mutton is steadily increased in China in recent years, which drives the development of the sheep raising industry. The sheep raising is changed from the original scattered raising of farmers to large-scale raising. In the production of the sheep, the quality of the stud ram determines the quality of the offspring of the sheep flock and determines the economic benefit of the sheep breeding. The artificial insemination technology can effectively promote the rapid propagation of the excellent variety, wherein the combination of the sheep semen diluent powder and the artificial insemination technology has very wide market prospect in the modern sheep raising industry. Firstly, the artificial insemination technology can effectively block the infection of reproductive diseases caused by natural mating; secondly, the artificial insemination technology can not only make full use of the genetic resources of the ram of the excellent breed, but also reduce the breeding cost by reducing the number of the ram of the breeding breed; moreover, the semen can be stored after being diluted, thereby being beneficial to long-distance transportation and overcoming geographical intervals. The diluent plays roles of diluting, buffering, nourishing and protecting sperms in the diluting and storing processes, and can effectively reduce the influence of low temperature on the sperms.
Disclosure of Invention
In order to solve the defects in the prior art, the invention aims to provide sheep semen diluent powder which is in a powder form and is used as ready-to-use, all components are matched with each other, so that the sheep semen diluent powder has a good activity enhancing effect on sheep semen, enhances the anti-stress property of sheep semen, particularly has a strong anti-oxidation and anti-bacterial effect, can effectively reduce refrigeration damage, and improves the survival rate of sperms and the integrity rate of acrosomes, thereby improving the fertilization rate.
In order to achieve the purpose, the invention adopts the specific scheme that:
a sheep semen diluent powder is prepared by dissolving sheep semen diluent powder in PBS buffer solution to prepare sheep semen diluent; the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.1-0.3g of monoammonium glycyrrhizinate, 0.5-1.0g of glutamic acid, 0.5-1.0g of L-arginine, 0.5-1.0g of melatonin, 37-40g of glucose, 4-8g of citric acid, 1-2g of EDTA, 1-2g of sodium carbonate and 0.1-0.3g of origanum oil.
As a further optimization of the scheme, the pH value of the sheep semen diluent is 6.8.
As a further optimization of the scheme, the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.1g of monoammonium glycyrrhizinate, 1.0g of glutamic acid, 1.0g of L-arginine, 1.0g of melatonin, 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate and 0.1g of origanum oil.
As a further optimization of the scheme, the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.2g of monoammonium glycyrrhizinate, 1.0g of glutamic acid, 1.0g of L-arginine, 1.0g of melatonin, 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate and 0.2g of origanum oil.
As a further optimization of the scheme, the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.3g of monoammonium glycyrrhizinate, 0.5g of glutamic acid, 0.5g of L-arginine, 0.5g of melatonin, 37g of glucose, 4g of citric acid, 1g of EDTA, 1g of sodium carbonate and 0.3g of origanum oil.
Has the advantages that:
1. the sheep semen diluent powder is in a powder form, is easy to store, is prepared at present when used, and reduces the pollution risk. The components are matched with each other, so that the sheep semen cold storage liquid has a good activity enhancing effect on sheep semen, enhances the stress resistance of sheep semen, particularly has a strong effect on oxidation resistance, can effectively reduce cold storage damage, and improves the sperm survival rate and acrosome integrity rate, thereby improving the fertilization rate. In the aspect of anti-oxidative stress, the monoammonium glycyrrhizinate and the oregano oil can act synergistically to play an additive effect; the monoammonium glycyrrhizinate is mainly used for improving the activity of superoxide dismutase, so that the capacity of clearing free radicals is improved, and the integrity of a cell membrane structure is protected; meanwhile, the glycyrrhiza trimonium salt has a protective effect on chemically damaged cells; it also has protective effect on photoaging of cells. The origanum oil can directly remove free radicals and improve the oxidation resistance. The oregano oil contains various compounds, and the main oxidation effect is phenolic acids and terpenoids. The phenolic acid compound contains phenolic hydroxyl which is a group with antioxidant activity, and the antioxidant is mainly used for terminating chain reaction by generating relatively stable phenoxy free radical through dehydrogenation reaction in the process of scavenging the alkoxy free radical. The two components can scavenge free radicals through different channels, and have additive effect and better sperm protecting effect.
2. The sheep semen diluent powder adopts plant essential oil to replace the traditional antibiotics, so that the negative effect of bacterial reproduction on semen caused by drug resistance is prevented. The monoammonium glycyrrhizinate and the oregano oil have antioxidant and antibacterial effects, so that sperms can be protected, and the vitality and the storage time of the sperms can be improved. The main components of the origanum oil playing an antibacterial role are phenols and terpenes, the phenols can change the permeability of cell membranes by denaturing proteins in the cell membranes or react with phospholipids in the cell membranes to destroy the synthesis of the proteins, so that the growth of microbial cells is inhibited, and the terpenes can also influence the DNA self-replication process in bacteria, so that the propagation and growth processes of the bacteria are inhibited, and the growth of thalli is inhibited. The mono-ammonium glycyrrhizinate has a good inhibitory effect on virus pathogenic microorganisms. The mono-ammonium glycyrrhizinate can inhibit the activity of virus polymerase to inhibit virus replication; indirect viral replication by reducing the activity of protein kinase C; the potential for inhibiting virus replication by inducing beta-chemokine production, and the potential for inhibiting virus propagation and spread between cells by reducing cell membrane fluidity to reduce cell-cell fusion. The monoammonium glycyrrhizinate and the oregano oil can kill pathogenic microorganisms through different mechanisms, have additive effect and have better protection effect on sperms.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention.
Example 1
The sheep semen diluent powder comprises the following components in percentage by weight: 0.1g of monoammonium glycyrrhizinate, 1.0g of glutamic acid, 1.0g of L-arginine and 1.0g of melatonin; 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate and 0.1g of oregano oil, wherein the diluted powder of the formula and PBS buffer solution are prepared into 1L of diluent, and the pH value is adjusted to 6.8.
Example 2
The sheep semen diluent powder comprises the following components in percentage by weight: 0.2g of monoammonium glycyrrhizinate, 1.0g of glutamic acid, 1.0g of L-arginine and 1.0g of melatonin; 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate and 0.2g of oregano oil, wherein the diluted powder of the formula and PBS buffer solution are prepared into 1L of diluent, and the pH value is adjusted to 6.8.
Example 3
The sheep semen diluent powder comprises the following components in percentage by weight: 0.3g of monoammonium glycyrrhizinate, 0.5g of glutamic acid, 0.5g of L-arginine and 0.5g of melatonin; 37g of glucose, 4g of citric acid, 1g of EDTA, 1g of sodium carbonate and 0.3g of oregano oil, wherein the diluted powder of the formula and PBS buffer solution are prepared into 1L of diluent, and the pH value is adjusted to 6.8.
Comparative example 1 sheep semen dilution powder, the formulation composition is as follows: 1.0g of glutamic acid, 1.0g of L-arginine and 1.0g of melatonin; 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate and 0.2g of oregano oil, wherein the diluted powder of the formula and PBS buffer solution are prepared into 1L of diluent, and the pH value is adjusted to 6.8.
Comparative example 2 sheep semen dilution powder, the formulation composition is as follows: 1.0g of glutamic acid, 1.0g of L-arginine and 1.0g of melatonin; 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate, 100 ten thousand IU of penicillin and 100 ten thousand IU of streptomycin, wherein the diluent powder of the formula and PBS buffer solution are prepared into 1L of diluent, and the pH value is adjusted to 6.8.
The sperm cells of examples 1 to 3 and comparative examples 1 to 2 were subjected to cold storage (4 ℃ C.) for 5 days, and the sperm cell survival rates before and after the cold storage and the percentage of intact acrosomes were observed, and the results are shown in tables I and II.
TABLE-Change in sperm motility (%)
| Group of | Before refrigeration | Refrigerating for 1 day | Refrigerating for 2 days | Refrigerating for 5 days | Refrigerating for 7 days | Refrigerating for 9 days |
| Example 1 | 95.77±0.49 | 84.40±0.78 | 79.44±1.37 | 75.13±0.73 | 69.34±1.65 | 63.13±1.72 |
| Example 2 | 93.24±0.35 | 82.35±0.38 | 79.64±1.77 | 76.39±0.86 | 70.73±1.75 | 65.94±0.86 |
| Example 3 | 95.17±0.62 | 89.86±0.84 | 86.75±0.73 | 81.64±0.74 | 76.75±1.67 | 70.18±0.63 |
| Comparative example 1 | 94.23±0.34 | 81.47±0.92 | 74.89±1.54 | 65.13±0.36 | 61.49±1.59 | 50.42±0.33 |
| Comparative example 2 | 94.18±0.73 | 80.64±0.84 | 71.78±1.38 | 60.13±0.85 | 51.47±1.46 | 42.13±0.93 |
TABLE II sperm acrosome integrity change (%)
| Group of | Before refrigeration | Refrigerating for 1 day | Refrigerating for 2 days | Refrigerating for 5 days | Refrigerating for 7 days | Refrigerating for 9 days |
| Example 1 | 99.74±0.57 | 94.36±0.74 | 90.46±1.37 | 85.43±0.82 | 79.41±1.95 | 73.47±1.74 |
| Example 2 | 99.27±0.75 | 95.47±0.46 | 91.64±1.54 | 86.25±0.74 | 80.68±1.58 | 75.85±0.46 |
| Example 3 | 99.43±0.48 | 97.86±0.56 | 93.05±0.82 | 89.04±0.83 | 86.73±1.36 | 80.42±0.74 |
| Comparative example 1 | 99.62±0.46 | 91.44±0.83 | 84.89±1.42 | 75.53±0.63 | 71.76±1.73 | 60.18±0.56 |
| Comparative example 2 | 99.32±0.63 | 91.75±0.71 | 85.78±1.54 | 80.23±0.75 | 72.53±1.44 | 62.43±0.74 |
From the above table, it can be seen that the sperm motility rate and the sperm acrosome integrity rate of the examples 1-3 in the formula process of the invention are higher after the cryopreservation, especially the best example 3 is used, the motility rate after 9 days of the cryopreservation reaches 70.18, and the acrosome integrity rate reaches 80.42, which indicates that the formula A adopted by the invention has obvious effect. The effect is much worse when the preparation is carried out outside the range of the formula of the invention.
It should be noted that the above-mentioned embodiments illustrate rather than limit the scope of the invention, which is defined by the appended claims. It will be apparent to those skilled in the art that certain insubstantial modifications and adaptations of the present invention can be made without departing from the spirit and scope of the invention.
Claims (5)
1. A sheep semen diluent powder is prepared by dissolving sheep semen diluent powder in PBS buffer solution to prepare sheep semen diluent; the method is characterized in that: the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.1-0.3g of monoammonium glycyrrhizinate, 0.5-1.0g of glutamic acid, 0.5-1.0g of L-arginine, 0.5-1.0g of melatonin, 37-40g of glucose, 4-8g of citric acid, 1-2g of EDTA, 1-2g of sodium carbonate and 0.1-0.3g of origanum oil.
2. The diluted sheep semen solution as claimed in claim 1, which is characterized in that: the pH value of the sheep semen diluent is 6.8.
3. The diluted sheep semen solution as claimed in claim 1, which is characterized in that: the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.1g of monoammonium glycyrrhizinate, 1.0g of glutamic acid, 1.0g of L-arginine, 1.0g of melatonin, 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate and 0.1g of origanum oil.
4. The diluted sheep semen solution as claimed in claim 1, which is characterized in that: the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.2g of monoammonium glycyrrhizinate, 1.0g of glutamic acid, 1.0g of L-arginine, 1.0g of melatonin, 40g of glucose, 8g of citric acid, 2g of EDTA, 2g of sodium carbonate and 0.2g of origanum oil.
5. The diluted sheep semen solution as claimed in claim 1, which is characterized in that: the content of each component in the sheep semen diluent powder adopted by each 1L of sheep semen diluent is as follows: 0.3g of monoammonium glycyrrhizinate, 0.5g of glutamic acid, 0.5g of L-arginine, 0.5g of melatonin, 37g of glucose, 4g of citric acid, 1g of EDTA, 1g of sodium carbonate and 0.3g of origanum oil.
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| CN202010383699.XA CN111685102A (en) | 2020-05-08 | 2020-05-08 | Sheep sperm solution diluent powder |
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| CN202010383699.XA CN111685102A (en) | 2020-05-08 | 2020-05-08 | Sheep sperm solution diluent powder |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114009427A (en) * | 2021-12-11 | 2022-02-08 | 江苏省农业科学院 | Rabbit semen cryopreservation diluent, preparation method, rabbit semen cryopreservation method and application |
| CN115245161A (en) * | 2022-08-17 | 2022-10-28 | 中国农业大学 | Oregano oil-based long-acting diluent for normal-temperature preservation of boar semen |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107232180A (en) * | 2017-05-25 | 2017-10-10 | 中国热带农业科学院热带作物品种资源研究所 | A kind of goat sperm dilution, preparation and its application process for being used to alleviate Heat-temperature stress |
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2020
- 2020-05-08 CN CN202010383699.XA patent/CN111685102A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107232180A (en) * | 2017-05-25 | 2017-10-10 | 中国热带农业科学院热带作物品种资源研究所 | A kind of goat sperm dilution, preparation and its application process for being used to alleviate Heat-temperature stress |
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| 何兰花: "新型植物抗生素-牛至油", 《畜牧与兽医》 * |
| 李方舟 等: "褪黑素对羊精液低温保存效果的影响", 《家畜生态学报》 * |
| 胡建宏 等: "牛冷冻精液稀释液中草药配方的初步研究", 《中国奶牛》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114009427A (en) * | 2021-12-11 | 2022-02-08 | 江苏省农业科学院 | Rabbit semen cryopreservation diluent, preparation method, rabbit semen cryopreservation method and application |
| CN115245161A (en) * | 2022-08-17 | 2022-10-28 | 中国农业大学 | Oregano oil-based long-acting diluent for normal-temperature preservation of boar semen |
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