CN119326065B - Application of Radix Isatidis and Green Vegetables in Improving Animal Semen Quality - Google Patents
Application of Radix Isatidis and Green Vegetables in Improving Animal Semen QualityInfo
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- CN119326065B CN119326065B CN202411610150.4A CN202411610150A CN119326065B CN 119326065 B CN119326065 B CN 119326065B CN 202411610150 A CN202411610150 A CN 202411610150A CN 119326065 B CN119326065 B CN 119326065B
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- 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
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/50—Feeding-stuffs specially adapted for particular animals for rodents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
- A61K36/315—Isatis, e.g. Dyer's woad
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
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Abstract
The invention provides an application of radix isatidis and green vegetables in improving the semen quality of animals, which is characterized in that the semen quality and the weight of the mice can be obviously improved by adding radix isatidis and green vegetable freeze-dried powder into the basic ration of the mice for feeding. The research result provides a solid foundation and theoretical guidance for the development and utilization of the isatis root greengrocery in the fields of health food, feed and the like, can further improve the added value attribute of the isatis root greengrocery, and has great practical significance.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to application of radix isatidis and green vegetables in improving the quality of animal semen.
Background
With the acceleration of modern life rhythm and the aggravation of environmental pollution, the quality decline of male sperms becomes a non-negligible problem, and the poor quality of sperms may lead to the failure of the fertilized eggs to be successfully combined, increasing the risks of early abortion and abortion in audit. Sperm is required to undergo three processes of production, growth and maturation in vivo, the number of sperm is reduced due to various reasons, and the problem of partial or total sperm motility is an important factor for causing male sterility of human beings, so that the improvement of sperm quality is vital to male fertility health and overall health. In the breeding process of domestic animals, semen quality plays an important role in variety improvement and population quality improvement. For example, the semen quality of a bull can directly affect the conception rate and the breeding efficiency of the cow, and is also a key factor affecting the production efficiency. The excellent boar semen quality not only can improve the economic benefit of the artificial insemination station, but also is an important guarantee for the reproductive performance of the pig farm. Therefore, the search for methods that effectively enhance semen quality is of great practical importance.
Isatis root vegetable (Isatis tinctoria oil No. 1) is obtained by hybridization fusion of Isatis tinctoria and somatic cells of Brassica napus, and multiple generation screening. The rape is widely planted in the world, has wide sources and large yield, and has rich nutritive value. The rape has multiple functions and uses, the leaves and the flower stems of the rape can be used as vegetables, the seeds can squeeze oil, the rapeseed meal and the green feed can be used for animals to eat, and the rape flowers have ornamental value. Rape is used as an important excellent oil and green feed crop, the planting area of China is wide, the annual planting area exceeds 1 hundred million mu, the yield of rape seeds exceeds 1,000 ten thousand tons, and the area and the total yield are all in the first place in the world. The isatis indigotica fort is an important medicinal plant in China and has the effects of resisting bacteria, resisting endotoxin, resisting cancer, regulating immunity, promoting blood circulation to remove blood stasis and resisting viruses, the root of the isatis indigotica fort is called isatis root, the leaf of the isatis indigotica fort is called dyers woad leaf, and the source and the yield of the isatis indigotica fort are far lower than those of rape. The isatis root green vegetable is obtained through the successive multi-generation backcross of the somatic cell hybrid formed by the somatic cell hybridization of cabbage type rape and woad and the cabbage type rape, and the relevant gene of the woad antiviral active ingredient is introduced into the rape to obtain the novel functional vegetable with broad-spectrum antiviral effect. The plant form and growth habit of the plant are similar to those of rape, and the plant is rich in the effective components of Isatis tinctoria G monomer additional system, and contains 17 amino acids and 8 microelements such as selenium, calcium, zinc and the like.
The existing researches show that the isatis root green vegetable has obvious anti-influenza virus H5N6 activity, the inhibition effect is equivalent to that of the conventional anti-influenza virus drug oseltamivir (darfein), and the isatis root green vegetable is obviously compared with a single board Lan Genjiang, can improve the weight loss and lung tissue lesions caused by influenza virus infection of mice, and has obvious anti-influenza virus infection effect. In addition, the study shows that the Isatis indigotica fort has remarkable inhibition effect on SARS-CoV-2 virus, can reduce weight loss caused by SARS-CoV-2 infection in a mouse infection model, improves the survival rate of mice, has remarkable anti-novel coronavirus effect and reduces the effect of novel coronavirus on the lung tissue injury of mice. However, whether the isatis root green vegetable can improve the semen quality is used as a novel functional vegetable has not been reported yet.
Disclosure of Invention
In view of the above, the invention provides an application of Isatis root vegetable in improving the semen quality of animals.
According to the invention, through a mouse feeding experiment, the isatis root and green vegetable freeze-dried powder is added into a mouse feed, and after continuous feeding, a sperm quality detection experiment is carried out, so that the effect of improving the sperm motility, the number and the effective sperm count of the mouse and reducing the sperm deformity rate is obvious when the freeze-dried powder is added into the feed for feeding the mouse.
Further, in the specific embodiment, the addition amount of the isatis root and green vegetable freeze-dried powder is 5% -20% of the mass of the basic ration.
Further, in a specific embodiment, the continuous feeding time is 5 to 7 weeks.
Compared with the prior art, the invention has the beneficial effects that:
(1) According to the invention, researches show that the semen quality of the mice can be remarkably improved by adding the isatis root and green vegetable freeze-dried powder with the mass fraction of 5% -20% into basic ration and feeding for 5-7 weeks. Wherein, the sperm quantity is improved by 9.8% -53.7%, the sperm density is increased by 13.6% -57.1%, the sperm motility is improved by 4.5% -13.3%, the effective sperm quantity is improved by 20.5% -175.5%, and the sperm malformation rate is reduced by 15.8% -28.6%; in addition, the weight of the mice is obviously improved within 1-5 weeks while the semen quality is improved.
(2) The research result of the mice provides solid foundation and theoretical guidance for the development and utilization of the isatis root greens in the fields of health food, feed and the like, and further improves the added value attribute of the isatis root greens.
Drawings
FIG. 1 shows the effect of adding radix Isatidis herba Sonchi Oleracei lyophilized powder to the basic ration on the sperm amount and density of mice.
FIG. 2 shows the effect of adding radix Isatidis herba Sonchi Oleracei lyophilized powder to the basic ration on the sperm motility and effective number of mice.
FIG. 3 shows the effect of adding radix Isatidis and herba Brassicae campestris lyophilized powder to the basal diet on mouse sperm malformation rate.
FIG. 4 shows the effect of adding radix Isatidis and herba Brassicae campestris lyophilized powder to the weight of mice.
Detailed Description
The present invention will be described in further detail with reference to specific examples so as to more clearly understand the present invention by those skilled in the art.
Key test material sources and physicochemical parameters:
Experimental animals SPF grade 4 week old male mice were derived from Liaoning Long biotechnology Co., ltd.
The isatis root and green vegetable freeze-dried powder used in the experiment is provided by Hubei Xiangqiang ecological agriculture limited company.
The specific raw materials are not described in the invention, and the raw materials are existing substances and can be directly purchased from the market.
Example 1
The embodiment provides a method for improving the semen quality of mice by using isatis root and green vegetable freeze-dried powder, which comprises the following specific steps:
1. Design of experiment
The single factor experiment design is adopted, wherein 40 SPF-class male Liaoning long mice with the age of about 4 weeks are selected and randomly divided into 4 groups, 10 mice are fed to a control group, 5%,10% and 20% of isatis root and green vegetable freeze-dried powder are respectively added to the basic diet by the test group, and the experiment period is 7 weeks.
The first stage is the feeding stage before 2024.8.13. All 40 male mice tested were fed in the same conventional diet pattern for 7 days.
The second stage, 2024.8.13 to 2024.10.1, is the feeding period, and the 40 mice to be tested are randomly divided into 4 groups of 10 mice each. The control group is fed with basic ration, and the test group is fed with the isatis root and green vegetable freeze-dried powder with the concentration of 5%,10% and 20% respectively added into the basic ration.
The body weight of the mice is measured periodically in the first stage and the second stage respectively, the semen of the mice is collected, and key quality indexes are measured.
2. Feeding management
The test mice are fed in a single cage, and eat and drink water freely. The raising environment is at the temperature of 22-26 ℃, the relative humidity is 50% -60%, the artificial illumination brightness is respectively 12 hours, and the basic ration feed formula is shown in the following table 1:
TABLE 1 details of basal ration formulation for mice
Further, in order to understand semen quality of mice of a control group and experimental groups of different addition amounts of the isatis root and the green vegetable freeze-dried powder, the following test was also performed.
1. Testing of sperm count and Density in mice
The mice were rapidly sacrificed after neck removal, epididymis was separated on one side, weighed and washed, sheared and placed on a 37 ℃ hot bench for 3min, and 10 μl of sample-adding needle was used for semen inhalation measurement. And (3) dripping the diluted semen on a blood cell counting plate, covering a cover glass, and observing and counting under a 200-400 times microscope. The density was obtained by taking the counted number of sperm (X) into the following equation:
X/80X 400X 10X dilution X l000 = number of sperm per ml semen
As shown in the figure 1, the result shows that the effect of adding the isatis root and green vegetable freeze-dried powder into the basic ration on the sperm amount of mice is shown as a result, the sperm amount of the mice with the isatis root and green vegetable freeze-dried powder added in the basic ration of 5% and 10% is not obviously different from that of the control group after the mice are fed for 5 weeks, the sperm amount of the mice with the isatis root and green vegetable freeze-dried powder added in the isatis root and the green vegetable freeze-dried powder added in the control group is obviously higher than that of the mice with the isatis root and green vegetable freeze-dried powder added in the control group, and the sperm amount of all mice in the experimental group is obviously higher than that of the control group after the mice are fed for 7 weeks. B represents the influence of adding the isatis root and green vegetable freeze-dried powder on the sperm density of mice in the basic ration, and the result shows that after 5 weeks of feeding, the sperm density of a control group is not obviously different from that of all experimental groups, and after 7 weeks of feeding, the sperm density of the mice with the isatis root and green vegetable freeze-dried powder added amount of 10% and 20% in the basic ration is respectively increased by 40% and 57% compared with that of the control group. The results prove that the radix isatidis green vegetable can improve the sperm quantity and the sperm density of mice. The graph shows the results of the comparison of the significance of the difference between the control group and the experimental group, wherein the difference is significant when the significance level P is less than 0.05, and the difference is extremely significant when the significance level P is less than 0.01.
2. Testing of mouse sperm motility and effective number
The percentage of sperm motility of forward and non-forward movement was observed under a mirror. The calculation mode is that the number of sperm (number/density-necrospermia/malformation)/density of linear advancing movement is multiplied by 100 percent. 200-400 sperm were observed under a microscope (400 x), the red one was dead, the colorless one was alive, and the number of effective sperm was recorded.
As shown in figure 2, A shows the influence of the addition of the radix isatidis and the green vegetable freeze-dried powder to the sperm motility of mice, and the result shows that after 5 weeks of feeding, the sperm motility of the experimental group is respectively improved by 6.8%,10.1% and 11.5% compared with the control group, the difference is obvious, and after seven weeks of feeding, the sperm motility of the experimental group is respectively improved by 4.4%,5.8% and 13.3% compared with the control group. B represents the influence of the addition of the isatis root and green vegetable freeze-dried powder in the basic ration on the effective sperm count of the mice, and the result shows that after 5 weeks of feeding, the effective sperm count of a control group is not obviously different from that of all experimental groups, and after 7 weeks of feeding, the effective sperm count of the mice with the addition of the isatis root and green vegetable freeze-dried powder in the basic ration of 10% and 20% is obviously higher than that of the control group, and the effective sperm count of the mice is respectively increased by 92.9% and 175.5%. The results prove that the radix isatidis green vegetable can improve the sperm motility and the effective number of mice. The graph shows the results of the comparison of the significance of the difference between the control group and the experimental group, wherein the difference is significant when the significance level P is less than 0.05, and the difference is very significant when the significance level P is less than 0.01.
3. Testing of mouse sperm malformation rate
The prepared smear was placed under a microscope (400 times), 200 sperm in different fields were counted, and the percentage of teratospermia was determined as the rate of teratospermia = number of teratospermia/number of total sperm examined x 100%.
As shown in figure 3, after 5 weeks of feeding, the sperm malformation rate of the experimental group is reduced by 18.2%,29.9% and 24.6% respectively compared with the control group, the difference is obvious, after 7 weeks of feeding, the sperm malformation rate of the experimental group is also obviously reduced by 15.8%,12.3% and 28.6% respectively compared with the control group, and the effect that the radix isatidis green vegetable can reduce the sperm malformation rate is demonstrated. The graph shows the results of the comparison of the significance of the difference between the control group and the experimental group, wherein the difference is significant when the significance level P is less than 0.05, and the difference is very significant when the significance level P is less than 0.01.
4. Test of mouse body weight
The mice in each group were weighed on an empty stomach before and after the test and each monday for 5 weeks continuously, and the average weights of the mice in each group were calculated, and the weight changes of the mice during the test were counted, and the results are shown in fig. 4, in which the weights of the mice in the control group and the mice in each test group were increased to different degrees. Specifically:
after one week of feeding, the weight of the mice in the control group is increased by 3.48g, the adding amount of the isatis root and the green vegetable freeze-dried powder in the basic ration is 5%, the weight of the mice with 20% is increased by 4.09g and 4g respectively without obvious difference, and the weight of the mice with the adding amount of 10% is increased by 5.36g, and the difference is obvious compared with the control group.
After two weeks of feeding, the weight of the mice in the control group increased by 8.15g, the adding amount of the isatis root and green vegetable freeze-dried powder in the basic ration was 5%, the weight of the mice with 20% increased by 8.84g and 8.1g respectively, and the weight of the mice with 10% added increased by 10.32g, and the difference is obvious compared with the control group.
After three weeks of feeding, the weight of the mice in the control group increased by 10.59g, the weight of the mice with the addition amount of the isatis root and the green vegetable freeze-dried powder in the basic ration increased by 11.64g, compared with the control group, no difference is obvious, and the weight of the mice with the addition amounts of 10% and 20% increased by 13.93g and 12.05g respectively, compared with the control group, the difference is obvious.
After four weeks of feeding, the weight of the mice in the control group increased by 12.16g, the weight of the mice with the addition amount of the isatis root and the green vegetable freeze-dried powder in the basic ration increased by 12.66g, and the mice with the addition amounts of 10% and 20% increased by 13.63g and 13.2g respectively, and the difference is obvious compared with the control group.
Five weeks after feeding, the weight of the mice in the control group increased by 14.58g, the addition amount of the isatis root and vegetable freeze-dried powder in the basic ration was 5%, the weight of the mice with 20% had no significant difference compared with the control group, and the weight of the mice with 10% had increased by 15.25g, and the difference compared with the control group was significant.
The results show that the isatis root green vegetables have the effect of promoting weight gain of mice. The results of the difference significance comparison between the control and experimental groups are shown in the figure, which shows that the difference is significant at a significance level P < 0.05.
The test results show that the addition of the isatis root and green vegetable freeze-dried powder in the basic ration of the mice can obviously improve the sperm quantity, density, vitality and effective sperm quantity of the mice and reduce sperm malformation. The method provides a solid foundation and theoretical guidance for the development and utilization of the isatis root green vegetables in the fields of health-care foods, feeds and the like.
The foregoing is merely a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (5)
1. Application of radix Isatidis and herba Brassicae Capitatae in preparing feed for improving animal semen quality is provided.
2. The use according to claim 1, wherein the semen quality index comprises sperm count, sperm density, sperm motility, effective sperm count, and sperm cell malformation.
3. The method for improving semen quality of mice according to claim 1, wherein the semen quality of mice is improved by adding radix Isatidis vegetable lyophilized powder into the basic ration of mice and continuously feeding.
4. The use according to claim 3, wherein the addition of the isatis root and green vegetable freeze-dried powder is 5% -20% of the mass of the basic ration.
5. Use according to claim 3, characterized in that the continuous feeding time is 5-7 weeks.
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| CN110637087B (en) * | 2017-03-08 | 2024-01-16 | 南京农业大学 | An epigenetic approach to manipulate plant phenotypic plasticity traits |
| CN109392829B (en) * | 2018-10-09 | 2022-05-17 | 湖北省畜禽育种中心 | Raising method and feed for improving the quantity and quality of bull semen |
| CN111066970A (en) * | 2019-12-30 | 2020-04-28 | 湖南百宜饲料科技有限公司 | Feed additive for improving reproductive performance of ternary sows |
| CN113181229B (en) * | 2021-04-30 | 2021-10-29 | 华中农业大学 | Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 |
| CN113789662B (en) * | 2021-09-07 | 2025-04-01 | 南通帝沙纺织有限公司 | Preparation method of plant antibacterial health care fabric containing fruit vitamins |
| CN114890931A (en) * | 2022-04-20 | 2022-08-12 | 湖北第二师范学院 | Extraction method and application of indole-3-acetonitrile in isati oil |
| CN117413770A (en) * | 2023-11-23 | 2024-01-19 | 山西农业大学 | Breeding method of green vegetable material with woad genetic material |
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| Rapid Identification of Alien Chromosome Fragments and Tracing of Bioactive Compound Genes in Intergeneric Hybrid offspring Between Brassica napus and Isatis Indigotica Based on AMAC Method;Guo YH;《International Journal of Molecular Sciences》;20250321;第5卷(第26期);2091-2105 * |
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