CN112586438A - Method for evaluating fertility of sires - Google Patents
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- CN112586438A CN112586438A CN202011214399.5A CN202011214399A CN112586438A CN 112586438 A CN112586438 A CN 112586438A CN 202011214399 A CN202011214399 A CN 202011214399A CN 112586438 A CN112586438 A CN 112586438A
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- 238000000034 method Methods 0.000 title claims abstract description 25
- 230000035558 fertility Effects 0.000 title claims abstract description 17
- 210000000582 semen Anatomy 0.000 claims abstract description 67
- 210000001550 testis Anatomy 0.000 claims abstract description 16
- 201000010063 epididymitis Diseases 0.000 claims abstract description 15
- 244000144972 livestock Species 0.000 claims abstract description 15
- 210000004267 spermatic cord Anatomy 0.000 claims abstract description 13
- 238000009395 breeding Methods 0.000 claims abstract description 11
- 230000001488 breeding effect Effects 0.000 claims abstract description 11
- 210000000918 epididymis Anatomy 0.000 claims abstract description 10
- 241001465754 Metazoa Species 0.000 claims abstract description 9
- 238000011156 evaluation Methods 0.000 claims abstract description 9
- 230000002159 abnormal effect Effects 0.000 claims abstract description 6
- 238000013210 evaluation model Methods 0.000 claims abstract description 6
- 238000001514 detection method Methods 0.000 claims abstract description 5
- 230000000694 effects Effects 0.000 claims abstract description 5
- 238000000611 regression analysis Methods 0.000 claims abstract description 3
- 230000008722 morphological abnormality Effects 0.000 claims description 15
- 230000004899 motility Effects 0.000 claims description 10
- 230000000750 progressive effect Effects 0.000 claims description 9
- 230000005856 abnormality Effects 0.000 claims description 7
- 150000001875 compounds Chemical class 0.000 claims description 6
- 230000002381 testicular Effects 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 5
- 230000035899 viability Effects 0.000 claims description 5
- 210000001215 vagina Anatomy 0.000 claims description 4
- 238000002738 Giemsa staining Methods 0.000 claims description 3
- 238000005259 measurement Methods 0.000 claims description 3
- 230000001419 dependent effect Effects 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 231100000527 sperm abnormality Toxicity 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000007774 longterm Effects 0.000 abstract description 2
- 238000012423 maintenance Methods 0.000 abstract description 2
- 238000012544 monitoring process Methods 0.000 abstract description 2
- 241000283074 Equus asinus Species 0.000 description 12
- 238000002604 ultrasonography Methods 0.000 description 12
- 210000002149 gonad Anatomy 0.000 description 8
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- 238000005516 engineering process Methods 0.000 description 4
- 241000283086 Equidae Species 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 206010067802 Sperm granuloma Diseases 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
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- 230000003902 lesion Effects 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 238000013441 quality evaluation Methods 0.000 description 2
- 230000001850 reproductive effect Effects 0.000 description 2
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- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 206010039954 Seminal vesiculitis Diseases 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
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- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
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- 238000003975 animal breeding Methods 0.000 description 1
- 206010003883 azoospermia Diseases 0.000 description 1
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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
- 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
- A01K29/00—Other apparatus for animal husbandry
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Abstract
The invention relates to a method for evaluating the fertility of a male animal, which specifically comprises the following steps: step 1: collecting semen; step 2: evaluating semen; and step 3: ultrasonic detection; and 4, step 4: measuring the testicle circumference; and 5: taking the abnormal percentage of the sperm morphology, the testis circumference, the epididymis tail cross section and the spermatic cord long axis parameter as independent variables, and establishing an evaluation model of the semen quality by adopting a multivariate stepwise regression analysis method; step 6: and calculating by adopting an evaluation model of the semen quality, evaluating the advancing activity, the total sperm count and the effective total sperm count of the sperms of the adult sires, and evaluating the semen quality. The method can reduce the long-term monitoring of the semen to the maximum extent, and can obtain the evaluation data about the production efficiency and quality of the semen. Provides a simple and practical method for selecting potential high-fertility sires for producers. The method saves equipment purchase and maintenance cost, and is suitable for evaluating the semen quality of the male livestock and breeding male livestock in large, medium and small farms.
Description
Technical Field
The invention relates to the technical field of male animal fertility evaluation and excellent breed male animal breeding, in particular to a method for evaluating the semen producing capability and the semen quality of male animals by detecting parameters of gonads and accessory gonads by using imaging technologies such as ultrasonic waves and the like.
Background
With the popularization and application of artificial insemination and embryo technology in livestock species breeding and animal production, the importance and economic value of excellent breeding male livestock in the industry are increasing, for example, one excellent male bull can produce 20 ten thousand doses of thin tube semen annually, one excellent male pig can produce 2 ten thousand semen bottle annually, and one horse can produce 2 ten thousand semen bottle annually. Therefore, breeding stock fertility assessment is extremely important in animal production. However, current breeding sire fertility assessments need to be determined by examining semen quality after the sire enters sexual maturity. On one hand, the method needs equipment and technology for semen collection and semen quality analysis, so that the cultivation cost of excellent male livestock is increased, on the other hand, the breeding performance of the male livestock is difficult to judge in regions with laggard grazing or production conditions, so that the semen quality indexes of the male livestock cannot be detected in small and medium-sized farms, the breeding and the maximum utilization value of the high-quality male livestock are hindered, and the popularization and the application of the artificial insemination technology are restricted.
Fertility robustness assessment includes clinical examination of males, as well as collection and assessment of semen to determine fertility of males. Ultrasound examination of the testis, epididymis and accessory gonads has proven to be a valuable and non-invasive method of assessing the morphology and pathology of mammalian genitalia. Gnemi and Lefebvre (2010) suggest ultrasound examination of the reproductive tract of bulls with azoospermia, an increased percentage of morphological abnormalities in sperm, abnormal size and shape of the reproductive organs, and pain of unknown cause. Transrectal ultrasonography of the parasgonads of horses is a routine reproductive robustness test commonly used to assess male horses, ejaculatory abnormalities or ejaculatory dysfunction. Ultrasound examinations were also performed on horses suspected of having abdominal pain due to seminal vesiculitis or genitourinary lesions. Ultrasonic diagnosis is reported in the detection of bulls, and the echo of the testis of the bulls is diagnosed by the ultrasonic diagnosis to be used as a puberty prediction marker of the bulls, but the puberty prediction marker is not superior to the puberty prediction marker by taking the scrotum circumference as a detection index. And testicular ultrasound image pixel intensity may have a relationship with sperm quality, but it has not been confirmed that ultrasound examination is most mainly used to detect lesion features in the testis and scrotum in the fertility robustness assessment of bulls. Ultrasound examination to measure boar testicular diameter is often used to determine the extent of pubertal development to rank similarly aged boars. Ultrasound examination is a reliable tool for examining testicles and epididymis, sperm granuloma may affect sperm quality, and ultrasound examination can provide valuable information for diagnosing ram sperm granuloma.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a mathematical model established based on parameters of the gonads and the accessory gonads to evaluate the semen quality of the male livestock.
The invention also aims to provide the application of the evaluation method in breeding male livestock.
The invention provides a method for evaluating the fertility of a male animal, which specifically comprises the following steps:
step 1: collecting semen: collecting semen by artificial vagina semen collection;
step 2: and (3) semen evaluation: performing semen evaluation on the collected semen to obtain sperm density (SC), Total Sperm Count (TSC), Total Motility (TM), percentage of Progressive Motility (PM) and percentage of sperm morphological abnormality (PSA);
and step 3: ultrasonic detection: measuring the epididymal tail cross-sectional area (CSACE) and the spermatic cord Long Axis (LASC) of the male stock by using B ultrasonic;
and 4, step 4: measuring the testis circumference: measuring the testis circumference of the male livestock by using a measuring tape to obtain the Testis Circumference (TC);
and 5: establishing a semen quality evaluation model by using a multivariate stepwise regression analysis method by taking the sperm morphological abnormality Percentage (PSA), the Testicle Circumference (TC), the epididymis tail cross-sectional area (CSACE) and the spermatic cord major axis (LASC) parameters as independent variables;
step 6: and (3) calculating by adopting an evaluation model of the semen quality, evaluating the advancing activity (PM), the Total Sperm Count (TSC) and the effective total sperm count (FSC) of the sperm of the adult sire and evaluating the semen quality.
On the basis of the scheme, the step 2 specifically comprises the following steps: evaluating semen within 10 minutes after semen collection, filtering semen with sterile gauze, removing gel fraction, measuring gel-free volume (GFV) in a measuring cylinder, and measuring sperm density (SC) with a photometer; total sperm number per ejaculation (TSC) was calculated using sperm density x gel-free volume;
analyzing the percentage of Total Motility (TM) and Progressive Motility (PM) using a computer-assisted sperm analyzer; for assessment of percent of total viability (TM), progressive viability (PM), sperm were dropped onto the slides, observing five discrete regions per sample;
estimating percent morphological abnormality (PSA) of sperm using Giemsa staining; percent morphological abnormality (PSA) of sperm was assessed after at least 3 tests.
On the basis of the scheme, the sperm morphological abnormality comprises head, middle, tail and compound abnormalities, and the compound abnormalities are more than 2 morphological abnormalities.
On the basis of the scheme, the epididymal tail cross-sectional area (CSACE) and the spermatic cord Long Axis (LASC) in the step 3 and the Testicular Circumference (TC) in the step 4 are repeatedly measured for three times to obtain an average value.
On the basis of the scheme, the semen quality evaluation model is as follows:
PM＝72.332+0.428LASC-0.441PSA，R2＝0.477，P＝0.000；
TSC＝-169.929+8.728TC+0.253CSACE，R2＝0.355，P＝0.002；
FSC＝-206.645+8.788TC+0.258CSACE，R2＝0.397，P＝0.001。
wherein: wherein R is2Expressing the interpretation rate of independent variable to dependent variable, P expressing the significant PSA expressing the abnormal percentage of sperm morphology, TC expressing the testicle circumference, CSACE expressing the cross-sectional area of epididymis tail,LASC represents the spermatic cord long axis, PM represents the progressive motility of sperms, TSC represents the total sperm count, and FSC represents the effective total sperm count;
on the basis of the scheme, the semen quality in the step 6 comprises the following steps: good, off-spec, and pass but not good;
if marching type vitality (PM)>85% and Total Sperm Count (TSC)>175.0×108Or effective total sperm count (FSC)>130.0×108Indicating that the semen quality is good;
if marching type vitality (PM)<75% and Total Sperm Count (TSC)<150.0×108Or effective total sperm count (FSC)<100.0×108Indicating that the semen quality is unqualified;
if 75 percent<Advance vitality (PM)<85% and 150.0X 108<Total Sperm Count (TSC)<175.0×108Or 100.0 × 108<Effective Total sperm count (FSC)<130.0×108Indicating that the semen quality is qualified but not excellent;
the method for evaluating the fertility of the sires is applied to breeding sires.
The invention has the beneficial effects that:
the invention provides a brand-new method for evaluating the semen quality of the sires, which is objective, simple and accurate in evaluation and can accurately predict the daily output and the sperm motility of the sires by easily obtained gonad parameters. Long-term monitoring of semen is minimized and assessment data regarding sperm production efficiency and quality can be obtained. Provides a simple and practical method for selecting potential high-fertility sires for producers. The method saves equipment purchase and maintenance cost, and is suitable for evaluating the semen quality of the male livestock and breeding male livestock in large, medium and small farms.
Drawings
The invention has the following drawings:
FIG. 1a is a first B-mode ultrasound image of the epididymis of a donkey;
FIG. 1B is a B-ultrasonic image of the epididymis of the donkey;
FIG. 1c is a first B-mode ultrasound image of the donkey's spermatic cord;
FIG. 1d is a second ultrasonic B-mode image of the donkey's spermatic cord;
FIG. 2 is a schematic diagram showing the comparison between observed and predicted values of forward activity;
FIG. 3 is a schematic diagram showing comparison between observed and predicted values of total sperm count;
FIG. 4 is a schematic diagram showing comparison between observed and predicted values of the number of effective total sperm.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
1. Semen collection
Semen is collected from 20 healthy and mature Texas donkeys, the age is 3-8 years, and the weight is 350-412 kg. Taking semen twice a week by using an artificial vagina. The same feeding and management conditions, 5 kg of ryegrass and 1.5 kg of concentrate, including: corn, bean cake, wheat bran, free drinking water. Before collection, the artificial vagina is lubricated with sterile gel (Minitub), preheated to 45 ℃, and mating activity is induced by natural heat-producing Jane.
2. Semen evaluation
Semen characteristics were assessed within 10 minutes immediately after semen collection. The semen was filtered using sterile gauze, the gel fraction removed, the Gel Free Volume (GFV) measured in a measuring cylinder, and the sperm density (SC) measured using a photometer (SDM-6; Minituub), the Total Sperm Count (TSC) per ejaculation being calculated as sperm density x gel free volume.
The percentage of Total (TM) and Progressive (PM) motility was analyzed using a computer assisted sperm analyzer (CASA, AndroVison; Minituub; Germany). For the assessment of the percentage of total viability (TM) and progressive viability (PM), five discrete areas were observed per sample using 5 μ l sperm droplets on the slides.
Percent morphological abnormality (PSA) of sperm was estimated by Giemsa staining. A total of 7 μ l of seminal fluid was smeared onto the slides and fixed with formaldehyde, then stained with Giemsa. Sperm morphological abnormality Percentage (PSA) was assessed after testing at least 3 slides per donkey.
The sperm morphological abnormality is divided into head, middle, tail and compound abnormalities, and the compound abnormalities comprise more than 2 morphological abnormalities.
3. Ultrasonic testing
First, the epididymal tail cross-sectional area (CSACE) and the spermatic cord major axis (LASC) of the donkey were measured using B-ultrasound, and each organ measurement was repeated 3 times as shown in fig. 1a to 1 d. The examined digital images are all stored for further analysis. All examinations and measurements were performed by the same skilled technician.
Next, The Circumference (TC) of each donkey testis was measured 3 times with a tape measure, and the average was taken for analysis.
Taking the sperm morphological abnormal Percentage (PSA), the Testicular Circumference (TC), the epididymal tail cross-sectional area (CSACE) and the spermatic cord major axis (LASC) parameters of the 20 male donkeys as independent variables, and calculating the advancing motility (PM), the Total Sperm Count (TSC) and the effective total sperm count (FSC) of the sperms through an evaluation model of the sperm quality.
As follows:
PM＝72.332+0.428LASC-0.441PSA，R2＝0.477,P＝0.000；
TSC＝-169.929+8.728TC+0.253CSACE，R2＝0.355,P＝0.002；
FSC＝-206.645+8.788TC+0.258CSACE，R2＝0.397,P＝0.001。
taking each donkey as an individual, and predicting the PM, TSC and FSC of the donkey by model calculation according to the semen, gonad and accessory gonad parameters of the donkey.
And finally, checking the coincidence degree of the predicted value and the observed value.
As a result, the correlation coefficients PM (r is 0.864 and P is 0.000), TSC (r is 0.527 and P is 0.017) and FSC (r is 0.630 and P is 0.003) between the predicted value and the measured value indicate that the predicted value and the measured value are better matched.
In addition, the prediction accuracy of the PM and FSC models is higher than the TSC model.
Those not described in detail in this specification are within the skill of the art.
Claims (7)
1. A method for evaluating the fertility of a male animal is characterized by comprising the following steps:
step 1: collecting semen: collecting semen by artificial vagina semen collection;
step 2: and (3) semen evaluation: carrying out semen evaluation on the collected semen to obtain the sperm density, the total sperm number, the total motility, the percentage of the progressive motility and the percentage of the abnormal sperm morphology;
and step 3: ultrasonic detection: b ultrasonic is used for measuring the cross section of the epididymis tail and the long axis of the spermatic cord of the male livestock;
and 4, step 4: measuring the testis circumference: measuring the testis circumference of the male livestock by using a measuring tape to obtain the testis circumference;
and 5: taking the abnormal percentage of the sperm morphology, the testis circumference, the epididymis tail cross section and the spermatic cord long axis parameter as independent variables, and establishing an evaluation model of the semen quality by adopting a multivariate stepwise regression analysis method;
step 6: and calculating by adopting an evaluation model of the semen quality to evaluate the semen quality.
2. The method for evaluating the fertility of a male animal according to claim 1, wherein the step 2 comprises the following steps: after semen collection, the semen is evaluated within 10 minutes, sterile gauze is used for filtering the semen, a gel part is removed, the gel-free volume is measured in a measuring cylinder, and the density of the semen is measured by a photometer; the total number of sperm per ejaculation was calculated using sperm density times gel-free volume;
analyzing the percentage of total and progressive motility using a computer-assisted sperm analyzer; for the assessment of the percentage of total, progressive viability, sperm were dropped onto the slides and five discrete regions were observed per sample;
estimating percent of morphological abnormality of the sperms by a Giemsa staining method; the percentage of morphological abnormality of the sperm was assessed after at least 3 measurements.
3. The method of evaluating fertility of a sire of claim 1, wherein the sperm morphological abnormality comprises a head, middle, tail and compound abnormality, wherein the compound abnormality is more than 2 morphological abnormalities.
4. The method of evaluating fertility of a sire according to claim 1, wherein the epididymal tail cross-sectional area and the spermatic cord major axis of step 3 and the testicular circumference of step 4 are measured in triplicate and averaged.
5. The method of evaluating fertility of a sire as in claim 1, wherein said model for assessment of semen quality is as follows:
PM＝72.332+0.428LASC-0.441PSA，R2＝0.477,P＝0.000；
TSC＝-169.929+8.728TC+0.253CSACE，R2＝0.355,P＝0.002；
FSC＝-206.645+8.788TC+0.258CSACE，R2＝0.397,P＝0.001；
wherein: wherein R is2The interpretation rate of independent variables to dependent variables is shown, P shows the significance of integral F test of a regression model, PSA shows the abnormal percentage of sperm morphology, TC shows the testicular circumference, CSACE shows the cross-sectional area of the epididymis tail, LASC shows the spermatic cord long axis, PM shows the advancing activity of the sperms, TSC shows the total sperm number, and FSC shows the effective total sperm number.
6. The method of evaluating fertility of a male animal according to claim 5, wherein the semen quality of step 6 comprises: good, off-spec, and pass but not good;
if the movement is advanced>85% and total sperm count>175.0×108Or effective total sperm count>130.0×108Indicating that the semen quality is good;
if the movement is advanced<75% and total sperm count<150.0×108Or effective total sperm count<100.0×108Indicating that the semen quality is unqualified;
if 75 percent<Advancing vitality<85% and 150.0X 108<Total sperm count<175.0×108Or 100.0 × 108<Effective total sperm count<130.0×108Indicating that the semen quality is acceptable but not good.
7. The use of the method for evaluating the semen quality of sires as defined in any one of claims 1-6 in breeding sires.
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