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1.
The ATP content in frozen and thawed semen from 17 bulls was determined by a bioluminescence method.The post thaw motility was assessed by phase contrast microscopy by subjective estimation of the percentage of sperm cells with forward motility. The concentration of spermatozoa was counted in a Bürker chamber. The correlation between ATP content and the number of sperm cells with forward motility was high.  相似文献   

2.
The effects of incubation temperature (+20 degrees C vs +35 degrees C) and media type on the ATP content and motility of spermatozoa were determined in fresh bovine semen in order to develop a method for assaying post-thaw quality. Semen was obtained from 3 bulls at 2 occasions. The spermatozoa were washed using a Ficoll-containing medium before being resuspended in each of 4 different media (I. 0.9% NaCl; II. Tris-buffer solution; III. seminal plasma; IV. seminal plasma+Tris-buffer solution) and incubated for 6 h. The least-squares means for ATP content were higher (p less than or equal to 0.05) at +20 degrees C than +35 degrees C for all media except no. I. By contrast, the least-squares means for sperm motility were higher (p less than or equal to 0.05) at +35 degrees C than at +20 degrees C in media II and III. A decrease over time in ATP content and motility at both temperatures was also observed. The single most important factor responsible for changes in ATP content and sperm motility was the temperature and the medium, respectively.  相似文献   

3.
This study was conducted to examine the effects of adding glutathione (1 mM) to media used for sperm washing and in vitro fertilization (IVF) on the improvement of early development of embryos produced using cryopreserved spermatozoa of the less IVF-competent bull (the one considered unqualified as spermatozoa supplier for the production of bovine blastocysts using IVF). The cryopreserved spermatozoa of this bull were characterized by normal motility and lower ATP content and blastocyst productivity than those of IVF-competent bulls. The addition of glutathione to the sperm washing medium was more effective in improving the productivity of blastocysts and ATP content than the addition of glutathione to the IVF medium or no glutathione addition at all (control). These results suggest that this simple method may be used to improve the potential of cryopreserved spermatozoa of less IVF-competent bulls to fertilize oocytes in vitro and to induce normal embryonic development after fertilization.  相似文献   

4.
In the present study, the correlations among age, body weight, scrotal circumference (SC), semen quality and peripheral testosterone and estradiol-17beta (E(2)) concentrations were investigated in pubertal (n=5) and postpubertal (n=7) groups of Holstein bulls over a 6 week period. There were significant positive correlations (P<0.01) among age, body weight and SC in both groups, and similar significant correlations between sperm motility and SC in pubertal bulls (P<0.01) and between sperm concentration and SC in postpubertal bulls (P<0.05). The sperm motility after collection (P<0.05) and after freezing and thawing (P<0.01) of the postpubertal bulls correlated positively with the E(2) concentration. Estrogen may be important for the function of postpubertal bull testes, in which it may regulate spermatozoa motility in vivo.  相似文献   

5.
Subjective microscopic sperm motility results have recently been demonstrated to differ between Holstein-Friesian (HF) and Belgian Blue (BB) bulls. However, such assessments are rather imprecise. In the present study, sperm motility was assessed objectively by means of the Hamilton Thorne CEROS version 12.2c computer-assisted sperm motility analyser (CASA), and differences between the BB and HF breed could also be demonstrated. Higher percentages of both totally (p < 0.0001) and progressively (p < 0.0001) motile spermatozoa were encountered in the HF breed compared with the BB breed. Furthermore, a lower kinetic efficiency of the BB spermatozoa, evidenced by a lower beat cross-frequency (p = 0.0007) combined with a higher lateral head displacement (p = 0.0015), was the basis for the lower velocity of BB sperm cells. Additionally, BB spermatozoa move less straight forward, resulting in a lower straightness (p < 0.0001). No sperm motility differences were observed between age groups within the BB breed. The breed differences were observed in the examined bull populations residing at AI centres, in Belgium for the BB bulls and in the Netherlands for the HF bulls. However, these bull populations are selected for fertility. A similar pattern was observed in an unselected bull population of both breeds, although these differences were mostly non-significant for the different CASA parameters. Nevertheless, these data suggest that a genetic component might be responsible for the observed sperm motility breed differences.  相似文献   

6.
The knobbed acrosome defect was found at levels of 25 to 100 percent of spermatozoa from 16 of 2054 beef bulls. The incidence of this defect appeared to be particularly high in the Charolais breed. Pedigree analysis of some of the affected Charolais bulls indicated there may be a genetic predisposition for this sperm defect. In eosin-nigrosin stained semen smears the most common form of the abnormality was a flattened or indented apex of the sperm head. A refractile bead at the apex of the sperm head was seen less commonly. Electron microscopy of the spermatozoa from one bull showed that the abnormality was similar to the knobbed sperm defect previously described in Friesian bulls. A breeding trial confirmed that bulls producing spermatozoa with a high incidence of knobbed acrosomes are infertile.  相似文献   

7.
Three experiments evaluated the effects of dietary Se and vitamin E on the ultrastructure of spermatozoa, ATP concentration of spermatozoa, and the effects of adding sodium selenite to semen extenders on subsequent sperm motility. The experiment was a 2 x 2 arrangement of treatments in a randomized complete block design. A total of 10 mature boars were fed from weaning to 18 mo of age diets fortified with two levels of supplemental Se (0 or .5 ppm) or vitamin E (0 or 220 IU/kg diet). The nonfortified diets contained .06 ppm Se and 4.4 IU vitamin E/kg. In Exp. 1, the spermatozoa from all boars were examined by electron microscopy. Vitamin E had no effect on structural abnormalities in the spermatozoa. When the low-Se diet was fed the acrosome or nuclei of the spermatozoa was unaffected, but the mitochondria in the tail midpiece were more oval with wider gaps between organelles. The plasma membrane connection to the tail midpiece was not tightly bound as when boars were fed Se. Immature spermatozoa with cytoplasmic droplets were more numerous when boars were fed the low-Se diet, but the occurrence of midpiece abnormalities occurred in boars fed diets with or without Se or vitamin E. Our results suggest that Se may enhance spermatozoa maturation in the epididymis and may reduce the number of sperm with cytoplasmic droplets. In Exp. 2, the concentration of ATP in the spermatozoa was evaluated in the semen of all treatment boars. When the low-Se diet was fed, ATP concentration was lower (P < .01), whereas vitamin E had no effect on ATP concentration. Experiment 3 investigated the effect of diluting boar semen with a semen extender with sodium selenite added at 0, .3, .6, or .9 ppm Se. Three ejaculates from each boar were used to evaluate these effects on sperm motility to 48 h after dilution. Sperm motility declined (P < .01) when Se was added to the extender, and this decline was exacerbated as the concentration of added Se increased (P < .01). The added Se was demonstrated to be tightly adhered to the spermatozoa. Overall, these results suggest that low Se-diets fed to boars resulted in abnormal spermatozoal mitochondria, a lower ATP concentration in the spermatozoa, and a loose apposition of the plasma membrane to the helical coil of the tail midpiece, but no effect from inadequate vitamin E was demonstrated. Adding sodium selenite to the semen extender reduced sperm cell motility.  相似文献   

8.
A study was conducted to evaluate the relationship between boar sperm motility and membrane integrity following exposure to media with 150–1120 mOsm. Total sperm motility was defined as the percentage of spermatozoa that had any form of motility was subjectively assessed under a light microscope. Sperm cell damage was expressed as a loss of membrane integrity as measured by a combination of fluorescent stains, carboxyfluorescein diacetate (CFDA) and propidium iodide (PI), and Hoechst 33258 (H33258). There were no significant differences between sperm motility and membrane-intact spermatozoa, as measured by CFDA-PI and H33258, in media with 250 and 300 mOsm. In anisosmotic conditions, a higher amount of membrane-intact spermatozoa than motile spermatozoa was observed. In hypo-osmotic conditions (150 mOsm), a high proportion of spermatozoa had curled or coiled tails and most of them retained their entire membrane integrity, as detected by CFDA-PI. In media with 350–1120 mOsm, some spermatozoa accumulated PI in the head region and CFDA in the mid-piece. These spermatozoa fluoresced blue at the lower region of the head, as detected by H33258. The ATP content in spermatozoa exposed to hypo- and hyperosmotic conditions was markedly reduced. There was no recovery of sperm motility on returning the spermatozoa to isosmotic conditions after 10 min incubation in anisosmotic conditions, indicating that the spermatozoa suffered an almost complete and irreversible loss of motility. This irreversible loss of motility may be a consequence of reduced ATP production in spermatozoa subjected to anisosmotic conditions. The results of this study demonstrate that plasma membrane integrity assessment in combination with sperm motility, using a range of media varying in osmolality, can give valuable information about the status and function of different sperm membranes, which might be relevant for semen preservation.  相似文献   

9.
The objective of this study was to examine differences in platelet-activating factor [1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine; PAF] in spermatozoa between two lines of Angus beef cattle divergently selected for blood serum insulin-like growth factor I (IGF-I) concentration. Endogenous lipids were extracted from the spermatozoa and endogenous PAF content was determined by radioimmunoassay. The amount of PAF detected in spermatozoa obtained from high IGF-I bulls (n = 8) ranged from 0.145 to 3.571 pM/10(6) cells. The level of PAF extracted from spermatozoa obtained from low IGF-I- bulls (n = 5) ranged from 0.001 to 1.024 pM/10(6) cells. Polynomial regression analysis revealed a significant cubic relationship (R(2) = 0.374; F = 6.292; P < 0.05) between spermatozoa PAF content and blood serum IGF-I concentration. Spermatozoa-derived PAF levels (mean +/- SEM) were significantly higher (P < 0.05) in the high IGF-I group (1.90 +/- 0.39 pM/10(6) cells) than in the low IGF-I group (0.59 +/- 0.20 pM/10(6) cells). High IGF-I bulls have a greater than three-fold higher PAF content in their spermatozoa than low IGF-I bulls. The data demonstrate that not only is PAF present in bull spermatozoa but that levels are significantly higher in individuals with high serum IGF-I concentrations.  相似文献   

10.
Epididymal spermatozoa from moose were studied in phase contrast, light interference and electron microscope. Some samples taken from cauda were diluted and frozen in liquid N2. The motility of the sperms after thawing was good.The concentration of spermatozoa in cauda was calculated to 10 × 106 cells per µl.Morphologically the spermatozoa of moose were found to be quite similar to those collected from bulls. The length of the sperm head was found to be approx. 8.8 µ and the average maximal width 5.2 µ. The average length of the tail was 54.7 µ and the entire length of the spermatozoon varied from 60 to 64 µ. Compared with sperm cells from bulls the moose spermatozoa appeared to have a somewhat shorter and broader head and a slightly shorter tail.The migration of the cytoplasmatic droplets, which was found to be completed in caput, seemed to follow the same pattern as in bulls and boars. As found in these species there was also in the moose a higher frequency of secondary abnormalities in the spermatozoa from cauda than in those from the other parts of epididymis.Studies of the fine structure of the moose spermatozoa seemed to indicate that these are of the same type as the spermatozoa of bulls, rams and boars. In sagittal sections the sperm head was thin, but in contrast to the sperm cells of the species mentioned above no typical waist-like narrowing in the equatorial region was found. The equatorial segment also seemed to be less arched than in the spermatozoa from bulls, rams and boars. Otherwise, no principal difference was found between ultrastructure of the moose spermatozoa and that of the spermatozoa collected from domestic species.  相似文献   

11.
Young beef bulls (n = 27) were used in a trial to study the effect of dihydrostreptomycin sulfate (DHS) or oxytetracycline (OTC) hydrochloride on spermatogenesis, epididymal sperm maturation, and freezability of sperm. Nine of the bulls were given a 22 mg/kg dose of DHS twice, 12 hours between doses. Nine other bulls were treated with OTC--1 dose of 26.4 mg/kg of body weight, and then 6 more doses each of 17.6 mg/kg, ca 12 hours between doses. The remaining 9 bulls were nontreated controls. The treatment regimens with the 2 antibiotics were without effect on spermatogenesis. These treatments also were without effect on seminal pH, ejaculate volume, percentage of motile spermatozoa, rate of spermatozoal motility, or concentration of spermatozoa in ejaculates harvested on day 3 or 7 of the study (day 0 = 1st day of treatment). There was a treatment-by-day effect on spermatozoal concentration; the number of sperm per milliliter was markedly increased on day 3 for OTC-treated bulls. The increased spermatozoal concentration in the OTC-treated group was associated with an influence of the antibiotic on ejaculation. All bulls given this antibiotic ejaculated without palpable penile engorgement or erection on day 3. On day 7 the rate of spermatozoal motility was increased in the 2 treatment groups compared with the rate in the control bulls. Also on day 7, the percentage of motile spermatozoa was greater in the OTC-treated bulls than in the control or DHS-treated bulls.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

12.
A QTL detection experiment was performed in French dairy cattle to search for QTL related to male fertility. Ten families, involving a total of 515 bulls, were phenotyped for ejaculated volume and sperm concentration, number of spermatozoa, motility, velocity, percentage of motile spermatozoa after thawing and abnormal spermatozoa. A set of 148 microsatellite markers were used to realize a genome scan. First, genetic parameters were estimated for all traits. Semen production traits were found to have moderate heritabilities (from 0.15 to 0.30) while some of the semen quality traits such as motility had high heritabilities (close to 0.60). Genetic correlations among traits showed negative relationships between volume and concentration and between volume and most quality traits such as motility or abnormal sperm while correlations between concentration and these traits were rather favourable. Percentages of abnormal sperm were negatively related to quality traits, especially with motility and velocity of spermatozoa. Three QTL related to abnormal sperm frequencies were significant at p < 0.01. In total, 11 QTL (p < 0.05) were detected. However, the number of QTL detected was within the range of expected false positives. Because of the lack of power to find QTL in this design further analyses are required to confirm these QTL.  相似文献   

13.
本试验采用PCR—RFLP方法分析了GnRH基因外显子2和LHR基因内含子9在144头中国荷斯坦牛和79头河南地方肉牛品种中的多态性,利用最小二乘法分析多态位点不同基因型与精液品质性状的关系。研究结果表明,2～3岁荷斯坦牛的鲜精活力显著高于4岁以上的牛,而畸形率显著低于7岁以上的牛。对2～4岁荷斯坦牛不同精液品质性状的简单相关分析表明,畸形率与顶体完整率和冻精活力呈显著的负相关（相关系数r分别为-0．736和-0．500）。不同基因型与精液品质性状的关联分析结果表明,144头中国荷斯坦牛所研究位点不同基因型对精液品质性状没有显著影响。而河南地方肉牛GnRH基因外显子2的A883G位点GG基因型的精子密度显著低于AA和AG基因型．LHR基因G51656T位点的TT基因型精子密度显著高于GT基因型,未检测到GG基因型。并且发现随着年龄的增长,种公牛的精液品质逐渐变差。GnRH和LHR基因可作为影u向肉牛精液品质性状的候选基因。  相似文献   

14.
The present study identified few potential proteins in the spermatozoa of buffalo bulls that can be used as an aid in fertility determination through comparative proteomics. The sperm proteome of high‐fertile buffalo bulls was compared with that of low‐fertile buffalo bulls using two‐dimensional difference gel electrophoresis (2D‐DIGE), and the differentially expressed proteins were identified through mass spectrometric method. The protein interaction network and the functional bioinformatics analysis of differentially expressed proteins were also carried out. In the spermatozoa of high‐fertile bulls, 10 proteins were found overexpressed and 15 proteins were underexpressed at the level of twofold or more (p ≤ 0.05). The proteins overexpressed in high‐fertile spermatozoa were PDZD8, GTF2F2, ZNF397, KIZ, LOH12CR1, ACRBP, PRSS37, CYP11B2, F13A1 and SPO11, whereas those overexpressed in low‐fertile spermatozoa were MT1A, ATP5F1, CS, TCRB, PRODH2, HARS, IDH3A, SRPK3, Uncharacterized protein C9orf9 homolog isoform X4, TUBB2B, GPR4, PMP2, CTSL1, TPPP2 and EGFL6. The differential expression ranged from 2.0‐ to 6.1‐fold between the two groups, where CYP11B2 was high abundant in high‐fertile spermatozoa and MT1A was highly abundant in low‐fertile spermatozoa. Most of the proteins overexpressed in low‐fertile spermatozoa were related to energy metabolism and capacitation factors, pointing out the possible role of pre‐mature capacitation and cryo‐damages in reducing the fertility of cryopreserved buffalo spermatozoa.  相似文献   

15.
The predictability of semen quality of mature sires from measurements at an early age is not well established. The aim of the present study was to determine age-dependent changes in the quality of bull semen from six Estonian Holstein (EHF) bulls, processed when the sires were 3, 5 and 7 years old. Fertility data such as 60-day non-return to oestrus rates (60d-NRRs) were available for 3-year-old bulls. From each batch, semen straws were analysed immediately after thawing [i.e. post-thaw (PT)] (controls) and after a swim-up (SU) procedure. The analyses comprised subjective and computerized measurements of sperm motility using computer-assisted sperm analysis (CASA) as well as estimations of sperm concentration, morphology and membrane integrity. There was a significant (p < 0.05) increase in the percentage of sperm motility (SU), membrane integrity (PT, SU) and normal tail and acrosome morphology (SU) with an increase in the age of the sires. The percentage of total motile spermatozoa PT measured by CASA correlated between 3- and 7-, and between 5- and 7-year-old bulls (p < 0.05). In addition, the proportion of head abnormalities tended to correlate between all three age groups both PT and after SU (p < 0.1). The sperm parameters correlating with fertility were average path velocity (VAP) (p < 0.001), total motility as measured by CASA (p < 0.01), linearly motile spermatozoa (p < 0.05) and CASA-assessed numbers of motile spermatozoa (p < 0.05), all after SU selection. The results showed that overall semen quality examined at 3 years of age is related to the semen parameters later in bulls' life. Moreover, CASA-assessed motility after SU seems to be a reliable marker for semen quality assessment as it shows correlation not only between the ages, but also to field fertility.  相似文献   

16.
Contents: Repeatability and heritability coefficients were estimated for ejaculate volume, mass movement, individual motility, sperm concentration, first and second post-freezing control of motility, number of doses frozen, total number of spermatozoa and total number of motile spermatozoa per ejaculate in the first ten ejaculates of 175 Swiss Holstein and 673 Simmental young bulls (200 pure Simmental, 473 Simmental x Red Holstein crosses). Data were analysed using the univariate individual animal model with effects of interval between consecutive collections, age of bull at collection, season of collection, breed group of Simmental bulls according to percentage of Red Holstein genes, bull (within breed group) and permanent environment. Crossbred bulls had a better performance in all traits. Repeatability coefficients ranged from 0.28 to 0.46. They indicate that the prediction of semen quality in young bulls of the Swiss Holstein and the Simmental breed is possible to some extent. Heritabilities in Swiss Holstein bulls were generally very low and did usually not exceed 0.10, but standard errors of these estimates were high. Relatively high heritabilities (0.18–0.30) were found for Simmental bulls, Differences in heritabilities between these two breed groups were probably a consequence of the low number of Swiss Holstein bulls.  相似文献   

17.
The spermiograms of 17 bulls were studied and 160 ejaculates were subjected to the thermoresistance test (38 degrees C) to evaluate sperm survival after thawing. After the first insemination of 10 682 cows, statistically significant differences were found in the fertilizing capacity of the ejaculates with various values of the thermo-resistance test. The best sperm fertilizing capacity was obtained in the ejaculates which retained progressive movement in 40% of the spermatozoa after two hours of exposure to the thermoresistance test. Out of the 1496 cows inseminated, 971 (i.e. 64.9%) got in calf, whereas after the insemination of 4216 cows with semen where only 30% of spermatozoa moved progressively at the end of the test, the number of pregnant dams was 2403, i.e. 56.98%; this difference is statistically significant (p0.05). At a lower sperm activity in the test the fertility after the first insemination was even lower. Although there was some difference in the individual fertility of bulls (54 to 67%), a positive relationship between the results of the thermoresistance test and fertility was recorded in all bulls.  相似文献   

18.
In mammalian spermatozoa, intracellular calcium plays a major role in sperm functions like motility and capacitation. Cryopreservation-induced modifications to sperm membrane result in an influx of intracellular calcium affecting calcium-dependent intracellular signalling pathways. Intracellular calcium activates adenyl cyclase to produce cAMP that activates phospholipase A(2) (PLA(2) ) and phospholipase C (PLC) generating lysophosphatidyl choline, 1,2-diacylglycerol (DAG) and IP(3) , acting as intracellular secondary messengers required for sperm capacitation. Present study was designed to determine levels of intracellular calcium, cAMP and DAG in fresh and frozen-thawed buffalo spermatozoa cryopreserved in the presence and absence of taurine or trehalose. A total number of nine ejaculates from three randomly chosen buffalo bulls were cryopreserved in Tris-based egg yolk extender and thawed in warm water at 37°C. The cAMP was measured by enzyme immuno assay, and intracellular calcium was quantified using fluorescent dye FURA 2-AM. Total lipid was extracted from spermatozoa, and DAG was estimated using thin layer chromatography followed by spectrophotometric analysis. Intracellular calcium, cAMP and DAG levels in spermatozoa were significantly (p < 0.01) increased following cryopreservation as compared to fresh ejaculate. Addition of taurine or trehalose to the freezing medium significantly decreased (p < 0.01) the levels of intracellular calcium and cAMP in frozen-thawed spermatozoa. 1,2-diacylglycerol content was also decreased significantly (p < 0.01) in spermatozoa cryopreserved in presence of additives. Moreover, significant (p < 0.01) improvement in post-thaw motility, viability and membrane integrity of spermatozoa on addition of taurine or trehalose clearly indicated the reduced level of capacitation-like changes in buffalo spermatozoa.  相似文献   

19.
Bull spermatozoa samples contain variable portion of motile and normal morphology spermatozoa along with spermatozoa incapable of fertilization due to their pathologic changes. As semen quality is influenced by biochemical and morphological characteristics of all spermatozoa, the aim of the study was to separate spermatozoa in discontinuous iodixanol density gradient solution and to determine their cholesterol, phospholipid, triacylglycerol and lipid peroxide concentrations and creatine kinase activity. The study was performed in winter and included seven Simmental bulls aged 1.5-3.5 years. Semen samples were collected by use of artificial vagina. Upon evaluation of semen quality (volume, concentration and progressive sperm motility), the samples were centrifuged in iodixanol density solution to obtain two sperm fractions. The two fractions included sperms with progressive motility greater than 90% and less than 20%, respectively. A statistically significantly higher lipid peroxide concentration was determined in sperm fraction with <20% progressive motility. Different sperm subpopulations can be obtained by separating bull spermatozoa in different iodixanol density gradient solutions, while monitoring their biochemical properties can help assess the sperm quality.  相似文献   

20.
This study aimed to develop a system of in vitro assays based on zona pellucida binding and in vitro fertilization for predicting male fertility in buffalo bulls. Frozen–thawed semen from nine bulls was tested for motility, viability index, acrosomal integrity, zona pellucida binding and in vitro fertilizing ability. Differences in post-thaw sperm motility between bulls were not significant. Differences in viability indices and percentage of spermatozoa with detached acrosome between bulls was highly significant (P < 0.001). Sperm attached per ovum, fertilization rates and polyspermy percentages varied significantly (P < 0.01) among buffalo bulls. A significant (P < 0.01) positive correlation coefficient of 0.69 was evident between normal acrosome and sperm attached per ovum, while between normal acrosome and fertilization efficiency it was 0.72. Sperm from different buffalo bulls differs in their ability to bind and fertilize oocytes. This study provides a basis to predict and maximize the in vitro fertilization performance of individual bulls.  相似文献   

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