Method for recovering sperm production of breeder cocks without sperm or with weak sperm

Abstract

The invention discloses a method for recovering sperm production of breeder cocks without / with weak sperm, and belongs to the technical field of poultry breeding. The method comprises the following steps: collecting seminal fluid of healthy cocks of more than 28 weeks old for two consecutive weeks, and selecting non-seminal breeder cocks; two weeks before fasting, the screened breeding cocks are transferred to a closed henhouse, and stress relieving medicine is added into drinking water; feeding and water supply are cut off from the first day to the third day from fasting, illumination is adjusted, and water supply is recovered from the fourth day; when the weight loss rate of the breeder cock group reaches 25-30%, gradually recovering feeding and illumination, and if the death rate in the implementation period exceeds 5%, stopping hunger, and recovering feeding and illumination; and observing the health condition of the chicken flocks every day, detecting whether the breeding cocks generate sperms or not 15 days after the breeding cocks recover to eat, and monitoring the body weight at the same time. The invention provides a method for recovering sperm production of breeder cocks without / with weak sperms, which can solve the problem that the hatching rate is influenced due to poor sperm quality or no sperm of the breeder cocks without / with weak sperms in the seed production.

Description

Technical Field

[0001] The invention belongs to the technical field of poultry breeding, and in particular relates to a method for restoring sperm production of sperm-free / weak sperm breed roosters. Background Art

[0002] In the production of chicken seeds, azoospermia is a common problem. About 10%-20% of breeding roosters will have azoospermia, which may be due to improper operation resulting in less or no semen, leading to asthenospermia, and then causing the loss of reproductive ability of the roosters. This situation not only reduces the fertilization rate of breeding eggs, but also greatly increases the breeding cost. In order to compensate for the impact of azoospermia / asthenospermia on production performance, it is often necessary to breed more roosters in production, but this further increases the breeding cost. At present, there are relatively few treatments for azoospermia / asthenospermia in breeding roosters, and the commonly used treatment method is to add drugs. Although this method can achieve certain results, it is costly and is not suitable for actual production. Therefore, improving the azoospermia of breeding roosters is of great significance for the economic cost control of the breeding industry and the smooth development of seed conservation work. Summary of the invention

[0003] The present invention aims to provide a method for restoring sperm production of roosters without / weak sperm, which can solve the problem of poor semen quality or no semen of roosters without / weak sperm in seed production, affecting the hatching rate.

[0004] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows: A method for restoring sperm production of sperm-deficient or weak sperm breeder roosters, comprising the following steps: S1. Selection of breeding roosters: Semen was collected from healthy roosters over 28 weeks of age for two consecutive weeks, and breeding roosters with semen volume, sperm motility and sperm density of 0 were selected; S2. Pre-fasting period: Two weeks before fasting, transfer the selected breeding roosters in step S1 to a closed chicken house with adjustable lighting. The relative humidity of the chicken house is 60%-75%. Natural ventilation is combined with negative pressure longitudinal ventilation. Stress relief drugs are added to drinking water. During this period, the feed intake of each chicken is counted. S3. Fasting period: Weigh the breeder roosters one day before fasting, cut off feed and water from the first to the third day of fasting, adjust the lighting, and keep the chicken house environment quiet. Water supply will be restored from the fourth day, and the body weight will be measured every three days to calculate the mortality rate; S4, Feeding recovery period: When the weight loss rate of the breeding rooster group reaches 25-30%, feeding and lighting are resumed, or when the mortality rate during the fasting period exceeds 5%, starvation is also ended and feeding and lighting are resumed; S5. Observe the health status of the flock every day, keep the chicken house environment quiet, and test whether the breeding roosters produce sperm 15 days after resuming feeding, and monitor their body weight.

[0005] Preferably, in step S2, the stress relief drug is one or more of γ-aminobutyric acid and vitamin mixed additives.

[0006] Preferably, the concentration of GABA is 100 mg / Kg.

[0007] Preferably, the vitamin mixed additive mainly includes vitamin A, vitamin D3 and the like.

[0008] Preferably, in step S3, food and water are cut off from the first day to the third day after fasting begins, and the light exposure time is adjusted to 8 h / d.

[0009] Preferably, in step S3, water supply is resumed on the 4th day, and the stress relief drug γ-aminobutyric acid is added to the drinking water, with 100 mg of γ-aminobutyric acid added per kilogram of water.

[0010] Preferably, in step S4, the recovery of feeding is specifically: the feed intake is gradually increased, the feed intake on the first day of recovery is 10-15% of the feed intake before fasting, and the feed intake is increased in equal gradients every day until the normal feed intake is restored and the animal can feed freely.

[0011] Preferably, in step S4, the lighting is specifically as follows: starting to resume feeding, the lighting is gradually increased synchronously, increasing the lighting by 0.5-1h every day from the 1st to the 3rd day, increasing the lighting by 1h every day from the 4th to the 8th day, and continuously illuminating for 16h every day after the 9th day.

[0012] Compared with the prior art, the present invention has the following advantages and technical effects: The present invention discloses a method for restoring sperm production of sperm-less / weak sperm breeder roosters, which uses starvation induction combined with reduced light to disrupt the biological clock of the reproductive system of the sperm-less / weak sperm breeder roosters, and then restores feeding and light exposure, while adding stress-relieving substances to drinking water to relieve stress caused by hunger, light, etc. The present invention can effectively stimulate the self-repair ability of the breeder roosters, promote testicular atrophy, and accumulate strength for subsequent reproductive system remodeling and functional recovery.

[0013] The present invention discloses a method for restoring sperm production of sperm-free / weak sperm breeder roosters. By gradually restoring the feeding process, the esophageal and intestinal damage that may be caused by the chickens suddenly feeding freely after fasting treatment is effectively avoided, and ammonia poisoning caused by excessive feeding is reduced. The method not only helps to repair and reshape the reproductive system of sperm-free breeder roosters, but also restores their sperm production ability, relieves the pressure of breeding farms, improves the fertilization efficiency of breeding eggs, reduces the number of parent generation updates, and is conducive to the long-term stable preservation of genetic resources.

[0014] During the recovery process, the reproductive performance of the breeder roosters is promoted by gradually increasing the light duration from 8 hours to 16 hours. This method of gradually increasing the light duration avoids the stress response caused by a sudden increase in the light duration, helps to better regulate the sleep time of the breeder roosters, and then regulates their metabolism, promotes the production of androgens, and ultimately affects the production of semen.

[0015] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a statistical chart of the changes in body weight of the unspermed breed roosters at different periods in Example 1; Figure 2 The statistical diagram of the testicle weight changes of the sperm-free roosters at different stages in Example 1; Figure 3 It is a statistical chart showing the changes in the semen volume of the sperm-free roosters at different stages in Example 1; Figure 4 It is a statistical chart of the changes in sperm motility of sperm-free roosters at different stages in Example 1; Figure 5 It is a statistical diagram of the change of sperm density of sperm-free roosters at different stages in Example 1; Figure 6 This is a statistical chart of the changes in testosterone (T) in the blood of sperm-free roosters at different stages in Example 1; Figure 7 This is a statistical chart of changes in luteinizing hormone (LH) in the blood of sperm-free roosters at different stages in Example 1; Figure 8 This is a statistical chart showing changes in follicle stimulating hormone (FSH) in the blood of sperm-free roosters at different stages in Example 1; Fig. 9 The figure is a statistical chart of the changes of testosterone (T) in the testicular tissue of the sperm-free roosters at different stages in Example 1; Fig.10 A statistical diagram of changes in luteinizing hormone (LH) in the testicular tissue of the sperm-free rooster at different stages in Example 1; Fig.11 This is a statistical chart showing changes in follicle stimulating hormone (FSH) in the testicular tissue of the sperm-free roosters at different stages in Example 1; Fig.12 These are paraffin sections of testicular tissue of the sperm-free rooster at different stages in Example 1, wherein: Fig.12 A in the figure is a testicular tissue section taken one day before fasting (F0). Fig.12 B is a testicular tissue section taken on the 39th day (R39) after resuming feeding. DETAILED DESCRIPTION

[0017] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0018] Unless otherwise defined, technical or scientific terms used in the present invention shall have the common meanings understood by one having ordinary skills in the field to which the present invention belongs.

[0019] In the present invention, unless otherwise specified, other test materials and instruments and equipment are conventional test materials in the art and can be purchased through commercial channels.

[0020] Example 1 S1. Selection of breeding roosters: Select healthy 28-week-old Yunong "H-series" breeding roosters of similar physical condition and from which semen cannot be collected by artificial massage. The artificial massage method is to train the breeding roosters for artificial semen collection for one week, and then collect semen artificially every 2 days. Roosters that cannot collect semen for three consecutive times are considered as spermless roosters; S2. Pre-fasting period: Two weeks before fasting, the selected breeding roosters in step S1 were transferred to a closed and adjustable lighting chicken house. The relative humidity of the chicken house was 60%-75%. Natural ventilation was combined with negative pressure longitudinal ventilation. Dry powder was fed and artificial feeding was carried out 4 times a day. Chicken manure was cleaned every 3 days and disinfected every 3 days. Routine epidemic prevention and management were carried out. γ-aminobutyric acid was added to drinking water to relieve stress. The concentration of γ-aminobutyric acid after dilution was 100 mg / kg. The average feed intake of the experimental chicken group was 120 g / hen. S3. Fasting period: The breeder roosters were weighed one day before fasting (F0), and three sperm-free breeder roosters were randomly selected for slaughter. Their testicles were weighed and collected. Feed and water were cut off from the first to the third day of fasting. The lighting time was adjusted to 8h / day, and the chicken house environment was kept quiet. Water supply was restored on the fourth day, and 100mg γ-aminobutyric acid was added per kilogram of water. Body weight was measured every three days, and the mortality rate was counted. The breeder roosters began to die around the 12th day, and the mortality rate was controlled within 5%. On the 15th day of fasting, the group weight loss rate was 25.42%. Fasting was stopped, feeding was resumed, and body weight was measured (F15).

[0021] S4, feeding recovery period: feed intake gradually increases. On the first day of feeding recovery, feed intake is 15% of the amount before fasting, and water supply and light increase by 0.5h / d. On the second day of feeding recovery, feed intake is 30% of the amount before fasting, and water supply and light increase by 0.5h / d. On the third day of feeding recovery, feed intake is 45% of the amount before fasting, and water supply and light increase by 1h / d. On the fourth day of feeding recovery, feed intake is 60% of the amount before fasting, and water supply and light increase by 1h / d. On the 5th day, the feed intake was 75% of the amount before the fasting period, and the water supply and light were increased by 1h / d. On the 6th day after the resumption of feeding, the feed intake was 90% of the amount before the fasting period, and the water supply and light were increased by 1h / d. On the 7th day after the resumption of feeding, the feed intake was 100% of the amount before the fasting period, and the water supply and light were increased by 1h / d. On the 8th day after the resumption of feeding, the feed intake was 100% of the amount before the fasting period, and the water supply and light time were 15h / d. On the 9th day after the resumption of feeding, the animals were allowed to eat freely, with water supply and light time of 16h / d. S5. Observe the health status of the flock every day and keep the chicken house quiet. After 15 days of resuming feeding, a few sperm-free roosters have produced pale white semen. At this time, monitor the weight of the chickens. On the 30th day of recovery, most of the sperm-free roosters can produce semen. At this time, monitor the weight of the chickens. Thereafter, monitor the weight of the sperm-free roosters every 3 days. On the 39th day of resuming feeding, all the sperm-free roosters have produced semen. At this time, not only are the test roosters weighed (R39), but also the semen volume, sperm density, and sperm motility of the test roosters are measured. At the same time, select 3 test roosters for slaughter, weigh and collect the testicles.

[0022] HE staining was performed on the testicular tissues one day before fasting (F0) and on the 39th day after resuming feeding (R39).

[0023] The test results are as follows: The body weights of the "H-series" roosters of Henan Agricultural University at different stages in Example 1 are as follows: Figure 1 As shown. Figure 1 It can be seen that the weight of the breeding roosters gradually decreased during the fasting period and returned to the F0 level on the 15th day after resuming feeding.

[0024] Testicular weight of F0 and R39 of Yunong "H" breed roosters, such as Figure 2 As shown. Figure 2 It can be seen that the testicle weight of breeding roosters is greater in R39 than in F0.

[0025] The semen volume, sperm density and sperm motility of sperm-free Henan Agricultural "H Series" roosters F0 and R39, such as Figure 3-Figure 5 As shown by Figure 3-Figure 5It can be seen that the sperm-free Yunong "H series" roosters cannot collect semen under artificial massage, that is, the semen volume, sperm density, and sperm motility are all 0 at F0, and the ejaculation volume is more than 0.2mL, the sperm motility is more than 50%, and the sperm density is 1.5X10 9 More than pcs / mL.

[0026] The contents of testosterone, luteinizing hormone, and follicle-stimulating hormone in the blood and tissues of sperm-free Henan Agricultural "H Series" roosters F0 and R39, such as Figure 6-Figure 11 As shown by Figure 6-Figure 11 It can be seen that the levels of testosterone, luteinizing hormone and follicle-stimulating hormone in the blood and tissues of R39 are higher than those in F0.

[0027] In this embodiment, paraffin sections of the testicles of sperm-free Henan Agricultural "H-line" roosters F0 and R39 were prepared and HE staining was performed. Fig.12 As shown. Fig.12 It can be seen that at F0, the lumen of the seminiferous tubules is larger, with fewer primary and secondary spermatocytes and spermatids, and fewer free sperm in the lumen. At R39, the number of sperm in the maturation process and free sperm is significantly increased compared with F0, and the lumen diameter is significantly reduced.

[0028] Example 2 The model of feeding control physiological remodeling in this example is the same as that in Example 1, except that the breeder rooster is a dwarf royal concubine.

[0029] Example 3 The model of feeding control physiological remodeling in this embodiment is the same as that in embodiment 1, except that the breeding rooster is a noble concubine.

[0030] Example 4 The model of feeding control physiological remodeling in this example is the same as that in Example 1, except that the breeder rooster is a Gushi chicken.

[0031] Example 5 The model of feeding control physiological remodeling in this example is the same as that in Example 1, except that the breeder roosters are of the A line.

[0032] Example 6 The model of feeding control physiological remodeling in this example is the same as that in Example 1, except that the breeder roosters are of the G line.

[0033] Example 7 The model of feeding control physiological remodeling in this example is the same as that in Example 1, except that the breeder roosters are of the M line.

[0034] Example 8 The model of feeding control physiological remodeling in this example is the same as that in Example 1, except that the breeder roosters are of the R line.

[0035] The sperm quality of the different breeds of roosters in Examples 2-8 above was tested before and after fasting. The results are shown in Table 1.

[0036] Table 1 Sperm quality of different breeds of roosters before and after fasting induction

[0037] As shown in Table 1, the 7 roosters in Table 1 were all trained for semen collection for two consecutive weeks (semen volume, sperm motility and sperm density were 0). The semen volume, sperm motility and sperm density were evaluated one day before fasting (F0), and the indicators were all 0. On the 39th day after resuming feeding (R39), the ejaculation volume (≥0.2mL), sperm motility (≥50%) and sperm density (≥1.5X109 / mL) were improved.

[0038] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. A method for restoring sperm production of sperm-deficient / weak sperm roosters, characterized in that: The steps include: S1. Selection of breeding roosters: Semen was collected from healthy roosters over 28 weeks of age for two consecutive weeks, and breeding roosters with semen volume, sperm motility and sperm density of 0 were selected; S2. Pre-fasting period: Two weeks before fasting, transfer the selected breeding roosters in step S1 to a closed chicken house with adjustable lighting. The relative humidity of the chicken house is 60%-75%. Natural ventilation is combined with negative pressure longitudinal ventilation. Stress relief drugs are added to drinking water. During this period, the feed intake of each chicken is counted. S3. Fasting period: Weigh the breeder roosters one day before fasting, cut off feed and water from the first to the third day of fasting, adjust the lighting, and keep the chicken house environment quiet. Water supply will be restored from the fourth day, and the body weight will be measured every three days to calculate the mortality rate; S4, Feeding recovery period: When the weight loss rate of the breeding rooster group reaches 25-30%, feeding and lighting are resumed, or when the mortality rate during the fasting period exceeds 5%, starvation is also ended and feeding and lighting are resumed; S5. Observe the health status of the flock every day, keep the chicken house environment quiet, and test whether the breeding roosters produce sperm 15 days after resuming feeding, and monitor their body weight.

2. A method for restoring sperm production of a sperm-deficient or weak-sperm rooster according to claim 1, characterized in that: In step S2, the stress relief drug is one of γ-aminobutyric acid.

3. The method for restoring sperm production of sperm-deficient / weak sperm roosters according to claim 1, characterized in that: In step S3, food and water are cut off from the first day to the third day of fasting, and the light exposure time is adjusted to 8h / d.

4. The method for restoring sperm production of sperm-deficient / weak sperm roosters according to claim 1, characterized in that: In step S3, water supply is resumed on the fourth day, and the stress relief drug γ-aminobutyric acid is added to the drinking water, with 100 mg of γ-aminobutyric acid added per kilogram of water.

5. The method for restoring sperm production of sperm-deficient or weak sperm roosters according to claim 1, characterized in that: In step S4, the recovery of feeding specifically includes: gradually increasing the feed intake, the feed intake on the first day of recovery is 10-15% of the feed intake before fasting, and increasing it in equal gradients every day until the normal feed intake is restored and the patient can feed freely.

6. The method for restoring sperm production of sperm-deficient or weak sperm roosters according to claim 1, characterized in that: In step S4, the lighting is specifically as follows: starting to resume feeding, the lighting is gradually increased synchronously, increasing the lighting by 0.5-1h every day from the 1st to the 3rd day, increasing the lighting by 1h every day from the 4th to the 9th day, and continuously lighting for 16h every day after the 9th day.

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