CN112314592A - Egg chicken semen diluent and diluting method - Google Patents
Egg chicken semen diluent and diluting method Download PDFInfo
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- CN112314592A CN112314592A CN202011174197.2A CN202011174197A CN112314592A CN 112314592 A CN112314592 A CN 112314592A CN 202011174197 A CN202011174197 A CN 202011174197A CN 112314592 A CN112314592 A CN 112314592A
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- diluent
- semen
- inositol
- streptomycin
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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
- A01N1/0215—Disinfecting agents, e.g. antimicrobials for preserving living parts
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a hatching egg chicken semen diluent and a diluting method, wherein the diluent comprises the following components: sodium glutamate, glucose, inositol, potassium dihydrogen phosphate, magnesium sulfate tetrahydrate, penicillin, streptomycin and distilled water. According to the invention, by adding potassium dihydrogen phosphate and inositol, the sperm motility can be enhanced, the sperm survival time can be prolonged, and by adding penicillin and streptomycin, the bacterial pollution in the artificial insemination process can be effectively relieved, the problem of unstable use effect caused by the existing dilution formula is solved, so that the sperm motility, survival time, fertilization rate and emergence rate after dilution are improved.
Description
Technical Field
The invention relates to the technical field of poultry breeding, in particular to a hatching egg chicken semen diluent and a diluting method.
Background
The egg breeder cock has the advantages of less semen quantity, thick semen, low proportion of male and female, overlarge sperm density, over-intense competition, too short semen using time, large cock breeding amount, increased breeding cost and low fertilization rate if the semen is not diluted after being collected. Therefore, the collected semen needs to be diluted with a diluent before use.
The existing diluting formula mainly comprises glucose, trisodium citrate dihydrate, sodium glutamate, sodium acetate, salt and the like, and although the diluting solution can be diluted, the using effect is unstable (the sperm motility is poor, the survival time is short, the fertilization rate and the emergence rate are low), and the storage is inconvenient.
Disclosure of Invention
The invention aims to provide a hatching egg hen semen diluent and a diluting method thereof, which are used for improving the activity, survival time, fertilization rate and emergence rate of diluted sperms.
The invention is realized by the following technical scheme:
a laying hen semen diluent comprises the following components:
sodium glutamate, glucose, inositol, potassium dihydrogen phosphate, magnesium sulfate tetrahydrate, penicillin, streptomycin and distilled water.
Inositol has effects of promoting cell metabolism and promoting growth, is growth factor of animal and microorganism, and potassium dihydrogen phosphate can improve water retention capacity of cell and improve cytoplasmic concentration, and can effectively improve sperm activity and survival time when used together; penicillin and streptomycin are common antibiotics, can effectively relieve bacterial contamination by matching, can effectively prevent bacterial infection of diluent in the preparation process by adding penicillin and streptomycin in the formula, and can properly relieve bacterial infection caused by misoperation in the artificial insemination process.
In conclusion, the inositol and the monopotassium phosphate are added into the formula at the same time, and the synergistic effect of the inositol and the monopotassium phosphate can improve the sperm motility and the survival time, so that the semen is diluted by the diluent, the service time of the collected semen can be prolonged, and the subsequent fertilization rate and emergence rate can be improved.
Further, the paint comprises the following components in parts by weight:
28.5g of sodium glutamate, 5g of glucose, 2.5g of inositol, 0.65g of monopotassium phosphate, 0.7g of magnesium sulfate tetrahydrate, 0.1g of penicillin, 0.1g of streptomycin and 1000ml of distilled water.
Further, the paint consists of the following components in percentage by weight:
28.5g of sodium glutamate, 5g of glucose, 2.5g of inositol, 0.65g of monopotassium phosphate, 0.7g of magnesium sulfate tetrahydrate, 0.1g of penicillin, 0.1g of streptomycin and 1000ml of distilled water.
Further, the diluent is subpackaged in reagent bottles.
The split charging in the reagent bottle is not only beneficial to preservation, but also has the advantage of convenient use.
A method for diluting a hatching egg semen diluent comprises the following steps:
s1, dissolving sodium glutamate, glucose, inositol, potassium dihydrogen phosphate, magnesium sulfate tetrahydrate, penicillin and streptomycin in distilled water, subpackaging in reagent bottles, sterilizing and freezing for storage;
s2, taking the diluent out, thawing at normal temperature, and pre-heating to 30 ℃;
and S3, collecting semen according to a preset dilution ratio.
Further, the dilution process is as follows:
and (3) extracting the diluent by using a disposable syringe and slowly injecting the diluent into the semen, wherein the syringe slowly stirs in a clockwise direction while injecting the diluent until the semen and the diluent are uniformly mixed.
Further, the autoclave was sterilized for 1 hour.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. according to the invention, the inositol and the potassium dihydrogen phosphate are added into the formula at the same time, and the mutual synergistic effect of the inositol and the potassium dihydrogen phosphate can improve the sperm motility and survival time, so that the semen is diluted by the diluent, the service time of the collected semen can be prolonged, and the subsequent fertilization rate and emergence rate can be improved.
2. The invention can obviously improve the sperm motility and the survival time by reasonably designing the addition amount of the inositol and the monopotassium phosphate.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not used as limitations of the present invention.
Example 1:
the laying hen semen diluent consists of the following components in parts by weight:
28.5g of sodium glutamate, 5g of glucose, 2.5g of inositol, 0.65g of monopotassium phosphate, 0.7g of magnesium sulfate tetrahydrate, 0.1g of penicillin, 0.1g of streptomycin and 1000ml of distilled water.
Comparative example 1:
the laying hen semen diluent consists of the following components in parts by weight:
0.611g trisodium citrate dihydrate, 5g glucose, 0.876g sodium glutamate, 0.65g potassium dihydrogen phosphate, 12.7g dipotassium hydrogen phosphate, 0.1g penicillin, 0.1g streptomycin, 2.593g sodium acetate, 1.99g salt (NaCl), 1.03g Tris, 1000ml distilled water.
Comparative example 2:
the laying hen semen diluent consists of the following components in parts by weight:
14g trisodium citrate dihydrate, 10g glucose, 20g sodium glutamate, 3.6g potassium dihydrogen phosphate, 1000ml distilled water.
Comparative example 3:
a hatching egg chicken semen diluent is normal saline.
A method of diluting a dilution of egg hen semen as described in any one of examples 1-3, comprising the steps of:
s1, dissolving the components in distilled water according to a certain proportion, subpackaging in reagent bottles, sterilizing for 1 hour by adopting a sterilization autoclave, and freezing for storage;
s2, taking the diluent out, thawing at normal temperature, and pre-heating to 30 ℃;
s3, after the semen is collected, diluting the semen with a diluent according to the proportion of 1:
when in dilution, a 5ml disposable syringe is matched with a No. 12 needle head, the diluent is slowly injected into the semen, and the syringe is slowly stirred clockwise while injecting until the semen and the diluent are uniformly mixed, and the thawed diluent is used up in days.
To verify the technical effect of the present invention, the following tests were performed:
selecting 12000 sets of 250-day old roman pink breeding hens, collecting semen by a back massage method, immediately diluting and storing the collected semen by the formulas of the example 1-the example 3 and the comparative example 1-the comparative example 5 according to the method, and performing comparative analysis on sperm activity, survival time, fertilization rate and emergence rate:
1) and (3) measuring the sperm motility and survival time of the semen diluted by different diluents: and (3) evaluating sperm motility: an estimation method; survival time assessment: microscopically, a stopwatch was used (sperm dead all), and the results are shown in table 1:
TABLE 1
| Item | Example 1 | Comparative example 1 | Comparative example 2 | Comparative example 3 |
| Sperm motility | 16.95a | 15.60a | 10.85b | 11.75b |
| Time to live | 35.25a | 31.5ab | 27.5b | 28.5b |
2) Carrying out artificial insemination operation on chicken groups in groups to collect hatching eggs for hatching and determining: evaluation of fertilization rate: post-hatch data collection accounting, results are shown in Table 2
TABLE 2
| Experimental group | Number of eggs incubated | Fertilization Rate (%) | Rate of emergence (%) |
| Example 1 | 1680 | 95.11±0.46a | 43.1±0.75b |
| Comparative example 1 | 1680 | 93.92±0.8 | 42.6±1.16 |
| Comparative example 2 | 1680 | 92.87±1.51 | 41.4±1.85 |
| Comparative example 3 | 1680 | 93.22±2.04 | 41.7±1.67 |
| Control group | 1680 | 95.07±0.51a | 43.04+0.63b |
Note: when the marked letters are the same, the difference is not significant, (p >0.05), and when the marked letters are not the same, the difference is significant (p < 0.05).
Among them, the control was semen without dilution (original semen inseminating).
From the data in tables 1 and 2, it can be seen that:
1) according to the invention, by adding inositol and potassium dihydrogen phosphate in the formula, the sperm motility and survival time are both obviously improved, and the emergence rate and fertilization rate are also improved compared with the method of only adding potassium dihydrogen phosphate in the comparative example 1 and the comparative example 2.
) From the comparison of the data of examples 1 to 3 with comparative examples 4 to 5, it can be seen that: when the addition amount of inositol and potassium dihydrogen phosphate is out of the range of the present invention, the sperm motility and survival time improvement performance is relatively poor, and the emergence rate and fertilization rate are relatively reduced.
To verify the presence of potassium dihydrogen phosphate and inositol in the diluent to promote sperm motility, the observation was conducted by separately preparing a formulation containing potassium dihydrogen phosphate and inositol and a formulation without potassium dihydrogen phosphate and inositol, as shown in table 3:
| formulation A (self-contained) | Formulation B | |
| Medicine and food additive | Dosage (g) | Dosage (g) |
| Glucose | 5 | 5 |
| Glutamic acid sodium salt | 28.5 | 28.5 |
| Potassium dihydrogen phosphate | 0.65 | |
| Inositol | 2.5 | |
| 4 magnesium sulfate hydrate | 0.7 | 0.7 |
| Penicillin | 0.1 | 0.1 |
| Streptomycin | 0.1 | 0.1 |
| Distilled water | 1000(ml) | 1000(ml) |
Before the experiment, a plurality of breeder cocks are prepared, and diluent, a microscope, a glass slide, a cover glass and the like are prepared according to each formula.
And (2) uniformly mixing the collected semen, absorbing 2 ml of semen in each part, diluting the semen by using a corresponding diluent in a ratio of 1:1 for standby application, dripping 1 drop of the diluted semen on a glass slide, putting a cover glass, wherein the semen is preferably distributed in the space between the cover glass and the glass slide, and observing the semen by 400 times under a microscope (comparing according to proportion), wherein the linearly moving semen has fertilization capability, and the small-range circularly moving and slightly swinging semen does not have fertilization capability, has high density and high vitality and is distributed in a nebulous manner under the microscope.
The results are shown in Table 4 for 3 consecutive days, 3 times daily, 13:00, 15:00 and 17:00 respectively:
TABLE 4
| Item | Formulation A | Formulation B |
| Sperm motility | 16.87a | 11.53b |
| Time to live | 35.18a | 32.5b |
The observation experiment shows that the addition of the inositol and the monopotassium phosphate in the formula can effectively improve the sperm motility and the survival time, and particularly has outstanding performance in improving the sperm motility.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are merely exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (7)
1. The laying hen semen diluent is characterized by comprising the following components:
sodium glutamate, glucose, inositol, potassium dihydrogen phosphate, magnesium sulfate tetrahydrate, penicillin, streptomycin and distilled water.
2. The egg hen semen diluent as claimed in claim 1, comprising the following components by weight:
28.5g of sodium glutamate, 5g of glucose, 2.5g of inositol, 0.65g of monopotassium phosphate, 0.7g of magnesium sulfate tetrahydrate, 0.1g of penicillin, 0.1g of streptomycin and 1000ml of distilled water.
3. The egg hen semen diluent as claimed in claim 1, which consists of the following components by weight:
28.5g of sodium glutamate, 5g of glucose, 2.5g of inositol, 0.65g of monopotassium phosphate, 0.7g of magnesium sulfate tetrahydrate, 0.1g of penicillin, 0.1g of streptomycin and 1000ml of distilled water.
4. The egg hen semen diluent of claim 1, wherein the diluent is dispensed into a reagent bottle.
5. A method of diluting an egg hen semen diluent as claimed in any one of claims 1 to 4 comprising the steps of:
s1, dissolving sodium glutamate, glucose, inositol, potassium dihydrogen phosphate, magnesium sulfate tetrahydrate, penicillin and streptomycin in distilled water, subpackaging in reagent bottles, sterilizing and freezing for storage;
s2, taking the diluent out, thawing at normal temperature, and pre-heating to 30 ℃;
and S3, collecting semen according to a preset dilution ratio.
6. The method of claim 5, wherein the dilution process comprises:
and (3) extracting the diluent by using a disposable syringe and slowly injecting the diluent into the semen, wherein the syringe slowly stirs in a clockwise direction while injecting the diluent until the semen and the diluent are uniformly mixed.
7. The method of claim 5, wherein the autoclave is sterilized for 1 hour.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011174197.2A CN112314592A (en) | 2020-10-28 | 2020-10-28 | Egg chicken semen diluent and diluting method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011174197.2A CN112314592A (en) | 2020-10-28 | 2020-10-28 | Egg chicken semen diluent and diluting method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113615680A (en) * | 2021-08-20 | 2021-11-09 | 中国农业科学院北京畜牧兽医研究所 | Kit for chicken semen cryopreservation and artificial insemination and use method thereof |
| CN113973811A (en) * | 2021-11-26 | 2022-01-28 | 华中农业大学 | Muscovy duck semen diluent and application thereof |
| CN115443988A (en) * | 2022-10-14 | 2022-12-09 | 浙江省农业科学院 | Chicken semen cryopreservation diluent and application thereof |
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| CN102812950A (en) * | 2012-09-07 | 2012-12-12 | 江苏省家禽科学研究所 | Chicken semen diluent as well as preparation method and application thereof |
| CN104094926A (en) * | 2014-08-05 | 2014-10-15 | 燕海峰 | Chicken semen diluent and preparation and utilization methods thereof |
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2020
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| CN102812950A (en) * | 2012-09-07 | 2012-12-12 | 江苏省家禽科学研究所 | Chicken semen diluent as well as preparation method and application thereof |
| CN104094926A (en) * | 2014-08-05 | 2014-10-15 | 燕海峰 | Chicken semen diluent and preparation and utilization methods thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113615680A (en) * | 2021-08-20 | 2021-11-09 | 中国农业科学院北京畜牧兽医研究所 | Kit for chicken semen cryopreservation and artificial insemination and use method thereof |
| CN113973811A (en) * | 2021-11-26 | 2022-01-28 | 华中农业大学 | Muscovy duck semen diluent and application thereof |
| CN115443988A (en) * | 2022-10-14 | 2022-12-09 | 浙江省农业科学院 | Chicken semen cryopreservation diluent and application thereof |
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Address after: 621000 No.1 Leizu Avenue, Yanting County, Mianyang City, Sichuan Province Applicant after: Fengji Food Group Co.,Ltd. Address before: 551700 Qinglong sub district office, Jinhaihu New District, Bijie City, Guizhou Province Applicant before: Guizhou Fengji Ecological Agricultural Technology Co.,Ltd. |
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Application publication date: 20210205 |
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