Comparison of semen characteristics,sperm freezability and testosterone concentration between Duroc and Yorkshire boars during seasons

This study was carried out to investigate the effects of Duroc and Yorkshire boars and seasons influencing semen characteristics, frozen-thawed sperm viability and serum testosterone concentration. Yorkshire boars produced higher semen volume compared to Duroc boars among seasons. However, sperm concentration did not differ significantly between Duroc and Yorkshire boars among seasons. Sperm motility of frozen-thawed sperm in Yorkshire boars was higher than in Duroc boars in spring and summer. Normal acrosome of frozen-thawed sperm in Yorkshire boars was higher than in Duroc boars in winter. Serum testosterone concentrations in Duroc and Yorkshire boars were higher in spring than in summer, autumn and winter. Serum testosterone concentrations in spring and summer were higher in Yorkshire boars than in Duroc boars. In conclusion, when serum testosterone concentrations were higher in Duroc and Yorkshire boars among seasons, semen volume, sperm concentration and frozen-thawed sperm viability were higher.     

Effects of dietary L-carnitine supplementation on semen characteristics in boars

In some species, dietary supplementation with L-carnitine has been reported to increase sperm concentration and sperm motility. The objective of these experiments was to test the hypothesis that L-carnitine supplementation improves the semen characteristics of boars. In Experiment 1, boars (258 days of age) were fed daily a control diet (n = 9) or the control diet plus L-carnitine (500mg per day; n = 9 ). Semen was collected weekly from Weeks 0 to 15 and on 4 consecutive days during Week 16. Experiment 2 was similar to Experiment 1 except boars ( n = 10 per treatment) were 504 days of age. For the weekly and intensive collections there were no consistently positive effects of treatment on semen volume, sperm concentration, total spermatozoa, or sperm motility. Spermatozoa from L-carnitine-treated boars did not display an enhanced ability to maintain motility during 7-day liquid storage. In conclusion, indicators of semen quality were not enhanced by dietary supplementation of L-carnitine in boars.     

Early sexual maturity in local boars of Northeastern India: age-related changes in testicular growth, epididymal sperm characteristics and peripheral testosterone levels

The present study reports the age related changes in the peripheral testosterone levels, testicular and epididymal growth and development and cauda epididymal spermiogram in local pigs of Northeastern India, which attain sexual maturity around 3 months of age. Local boars (n = 20) were castrated at monthly intervals from 2 to 6 months of age (4 boars per month) to study the testicular growth and development and the epididymal spermiogram. Blood samples, collected from local boars (n = 6) at monthly intervals from 2 to 6 months of age, were analyzed for testosterone levels by radioimmunoassay. Compared to Hampshire boars, significantly (P < 0.05) high testosterone levels were observed in the local boars as early as 2 months of age. The mean (± SEM) level of testosterone in the local boars at 2, 3 and 4 months of age was 11.89 ± 1.52, 20.45 ± 1.33 and 20.38 ± 2.0 ngml⁻¹, respectively. Though there was consistently significant (P < 0.05) difference in the body weight between Hampshire and local pigs, the same was not observed in case of testicular weight except at 3 and 6 months of age. In line with the above observation, the testis:body weight ratio (gram testis per kg body weight) was significantly (P < 0.05) higher in the local boars compared to the Hampshire boars at any time of observation, which ranged from 0.8 to 1.0 in case of Hampshire and from 2.3 to 3.0 in local boars. The sperm concentration in the cauda epididymal fluid of local boars at 2, 3 and 6 months of age was 2255 ± 186.6, 3685 ± 103.8 and 4325 ± 146.2 million/ml, respectively and the sperm motility, viability and total abnormality was 73.3, 75.2 and 6.2%, respectively at 3 months of age. Taken together, the testosterone level, testicular growth and development and epididymal spermiogram indicate the trait of early sexual maturity in the local pigs as compared to Hampshire.     

Variation in testosterone production among boards and its relationship to sexual interest and breeding performance

Variation in ability to produce testosterone in response to both GnRH and ACTH administration and quatitative relationships between GnRH-stimulated testosterone levels, ACTH-stimulated testosterone levels, sexual interest and breeding performance were assessed in a group of 31 Duroc boars (115.4 +/- 2.5 kg body weight and 212.2 +/- 3.0 days of age). Mean area beneath the testosterone response curve increased (P<0.01) after GnRH and ACTH but the magnitude of response was variable among boars. Post-GnRH testosterone area varied from 7.44 to 50.86 ng/ml X h with a CV = 52.41% while post-ACTH testosterone area varied from 4.99 to 28.78 ng/ml X h with a CV = 45.46%. Mean sexual interest and mean breeding performance scores were correlated (r = 0.67, P<0.01); however, correlations of either variable with testosterone areas were low and nonsignificant. These results indicate that the testosterone-producing ability of boars of similar age and breeding is highly variable and suggest that peripheral testosterone concentrations may not be good indicators of either libido or breeding performance.     

Effects of a high semen-collection frequency on the quality of sperm from ejaculates and from six epididymal regions in boars

This study examines the effect of semen-collection rhythm on the sperm maturation process in boar epididymis. Three post-pubertal boars were submitted to a high semen-collection frequency (stressed boars) during 4 days, and three males were kept as a control group (control boars). Semen samples coming from six epididymal regions and from the ejaculate of each male were evaluated for sperm concentration, sperm vitality, sperm motility and sperm morphology. In each epididymal region, either fluid resorption or fluid secretion was determined from the variation in sperm concentration. The pattern of fluid resorption-secretion along the epididymal duct differed significantly between the stressed and control boars. A high semen-collection frequency also affected the development of sperm motility and the sperm cytoplasmic droplet displacement along the epididymal duct. The incidence of some sperm abnormalities was also found to be higher in some epididymal regions and ejaculates of stressed boars. From the results of this study, it can be concluded that a high semen-collection frequency brings about an altered resorption and secretion pattern of the epididymal fluid, which results in defective sperm maturation and abnormal development of sperm motility.     

Influence of supplemental lighting on serum LH,testosterone and semen quality in prepubertal and postpubertal boars

Thirty-six prepubertal boars received either natural lighting or supplemental lighting from incandescent or fluorescent sources between 8 and 35 weeks of age. Serum concentrations of luteinizing hormone (LH) and testosterone were similar among the treatment groups (P > 0.05). Even though libido scores were higher at 20–24 weeks of age in boars that received supplemental lighting, the libido scores in control boars and supplemental-lighted boars were similar by 30 weeks of age. From 44 to 83 weeks of age, 20 postpubertal boars were exposed to natural or supplemental lighting and semen was collected 3 times in one week per month. No differences in spermatozoa concentration, forward motility, or total motile spermatozoa were found among the treatments. Semen volume of boars maintained on supplemental lighting was lower (P < 0.05) than that of control boars switched to supplemental lighting and lower than that of boars exposed to supplemental lighting that were switched to control. No differences in serum LH or testosterone were observed among treatments in boars from 42 to 82 weeks of age. When boars were bilaterally castrated at 83 weeks of age, no differences in testes weight, epididymal weight, sperm per gram testes, relative testes weight, or relative epididymal weight were found. Data from the present experiment indicate that supplemental lighting does not enhance semen quality of postpubertal boars.     

慢性间歇性缺氧降低SD大鼠精子数量和活力的研究

摘要 目的：通过制备大鼠慢性间歇性缺氧的动物模型,观察间歇性缺氧对雄性大鼠精子数量和活力的影响,并初步探讨其可能的发生机制。方法：16只雄性SD大鼠随机分为对照组和缺氧组。间歇性缺氧8周后分别测定两组大鼠精子活力和精子浓度,睾丸组织中P53的表达量和细胞的凋亡水平,以及血清睾酮（T）、促卵泡素（FSH）和黄体生成素（LH）的浓度。结果：与对照组比较,缺氧组大鼠精子的浓度和活力均降低（P>0.05),并且前向运动的精子数明显减少（P<0.05）。同时,缺氧组睾丸组织P53 mRNA的表达和睾丸组织中凋亡的细胞均多于对照组,血清T、LH的浓度低于对照组（P<0.05）。结论：间歇性缺氧时可能影响了下丘脑-垂体-性腺轴,不仅使睾酮合成减少,还降低了对P53的抑制作用而诱导了大量细胞的凋亡,最终降低了SD大鼠的精子数量和活力。     

Characterization of the semen quality of postpuberal boars with spontaneous unilateral abdominal cryptorchidism on the right side.

In recent studies, we found that the ectopic testis from postpuberal boars with unilateral abdominal cryptorchidism does not produce sperm. Therefore, in these males, the seminal characteristics can be used as indicators of the activity of the scrotal testis and its epididymis and also the accessory glands. The semen quality (ejaculate volume, cell-rich fraction volume, sperm concentration, sperm vitality, sperm motility, sperm morphology and cephalic stability of spermatozoa) was evaluated in healthy postpuberal boars and in postpuberal boars with unilateral abdominal cryptorchidism on the right side. In comparison with the healthy boars, the unilateral abdominal cryptorchid boars showed a significant decrease of the ejaculate volume, sperm concentration and sperm motility. The low sperm concentration indicated that unilateral abdominal cryptorchidism severely impairs the sperm production of the scrotal testis. The decrease of ejaculate volume was attributed to an abnormal activity of the accessory glands. The alterations in sperm motility develop as a result of dysfunctions in the epididymal epithelium and/or the accessory glands. The sperm vitality, sperm morphology and cephalic stability of spermatozoa maintained normal values; therefore, at testicular level, despite the low sperm production, the germ cell differentiation is not disturbed. At epididymal level, the morphological maturation of spermatozoa is not altered.     

Seasonal variation in serum testosterone, testicular volume and semen characteristics in coatis (Nasua nasua)

The objective was to characterize seasonal changes in serum testosterone concentration, testicular volume and sperm quantity and quality in captive coatis (Nasua nasua) from Pantanal, MT, Brazil. Sampling was done once monthly for 1 y. Mean (± SEM) serum testosterone concentrations (767.37 ± 216.2 ng/ml) and total and progressive sperm motility (79.6 ± 3.9%; 3.8 ± 0.3, on a scale of 0 to 5) peaked in July. The highest combined testis volume (10.3 ± 0.4 cm³) and sperm concentration (403 million ± 102 sperm/ml) occurred in August, at the peak of the winter breeding season. No seasonal effects on percentages of morphologically normal sperm, acrosome integrity, or live sperm were detected; however, the percentage of secondary sperm defects was higher in the winter. In conclusion, intricate relationships between testosterone concentration, testis volume, semen concentration and total and progressive sperm motility with high levels of breeding activity were observed during the dry season in the winter (June, July, August), followed by a subsequent decline in these activities during the wet season (i.e., summer: December, January, February). There was no seasonal pattern for production of functionally intact and morphologically normal sperm.     

Semen changes in boars after experimental infection with porcine reproductive and respiratory syndrome (PRRS) virus

Eleven boars seronegative to porcine reproductive and respiratory syndrome virus (PRRSV) were trained for semen collection: five boars were inoculated intranasally with 6 x 10(6)TCID(50)/ml of PRRSV (Group A); four boars were inoculated intranasally with 6 x 10(4)TCID(50)/ml (Group B); and two boars were used as uninfected control (Group C). Semen samples were collected at 7-d intervals from 49 d prior to experimental inoculation with PRRSV to 70 d after inoculation, and were examined for sperm volume, sperm concentration, sperm morphology, sperm motility and for the presence of PRRSV. The infection in boars was demonstrated by the reisolation of PRRSV from the serum of all inoculated boars. Rectal temperatures and general health of the boars were clinically normal throughout the trial. Differences were observed in the quality of semen collected from boars after experimental infection with PRRSV. This infection induced a significant decrease in sperm motility and in spermatozoa with normal acrosomes. Of the semen samples tested for virus isolation in swine alveolar macrophages PRRSV was only isolated in 1 boar from Group B. The virus was detected in an additional semen sample in Group A by the production of an antibody titer in a biological assay. All attempts to detect PRRSV by RT-PCR in semen samples were unsuccessful. Nevertheless, from our study it is possible to suggest that the PRRSV can occasionally be transmitted in the semen during the initial phase of the disease.     

Aging-related reproductive decline in the male spider monkey (Ateles geoffroyi)

Background It is unknown whether male black‐handed spider monkeys (Ateles geoffroyi) undergo a reproductive decline as they age. The purpose of this work was investigating whether serum testosterone and semen quality decrease with age in these primates as occur in other species. Methods In this study, we assessed age variations in the concentration of serum testosterone, sperm counts, and motility in nine males. We took serum blood samples and semen samples by electroejaculation of anesthetized males throughout the fertile season. Results We found that testosterone levels and total sperm concentration, motile, progressive, fast linearly moving, medium linearly moving, slow linearly moving and immotile sperm significantly decreased with age. Conclusions Our results show that a reproductive decline occurs in male spider monkeys because of a decrease in sperm counts. These results are similar to those of rhesus monkeys and men, despite the phylogenetic distance between New and Old World primates.     

Seminal plasma damages sperm during cryopreservation, but its presence during thawing improves semen quality and conception rates in boars with poor post-thaw semen quality

To determine the effects of seminal plasma during and after cyopreservation on post-thaw sperm functions in semen from poor freezability boars, seminal plasma was removed immediately after collection, and sperm was subjected to cooling and freezing. Removal of seminal plasma did not significantly affect post-thaw sperm motility in good freezability boars; however, in boars with poor freezability, it increased post-thaw motility relative to control sperm cooled with seminal plasma (64.5+/-3.4% vs. 30.9+/-3.1%, P<0.01). Freezing sperm without seminal plasma increased both loss of the acrosome cap (37.5+/-1.6% vs. 18.4+/-2.8%, P<0.01) and expression of a 15 kDa tyrosine-phosphorylated protein (capacitation marker) in thawed sperm relative to controls; the addition of 10% (v/v) seminal plasma to the thawing solution significantly suppressed both changes and increased conception rate to AI (70% vs. 9% in the control group, P<0.05). In conclusion, our novel cryopreservation and thawing method increased the success of AI with frozen-thawed porcine semen, particularly from boars with poor post-thaw semen quality.     

The decline of porcine sperm motility by geldanamycin, a specific inhibitor of heat-shock protein 90 (HSP90)

Sperm motility is an important parameter for fertility. The molecular mechanisms of mammalian sperm motility are still largely undefined. Our previous observations suggested that heat shock protein 90 (HSP90) may be associated with porcine sperm motility. The aim of the present study was to further characterize the plausible novel function of HSP90 on sperm motility. Semen from normal, sexually mature boars with sperm motility higher than 80% was used. An HSP90-specific inhibitor, geldanamycin (GA), was added to diluted semen at 0.5, 1.0, 2.5 or 5.0 microg/mL and the semen was then incubated at 37 degrees C for 15, 30, 45 or 60 min. Sperm motility was determined by using computer-assisted semen analyzer at the end of incubation. The results indicated that GA significantly reduced sperm motility in a dose and time dependent manner. Moreover, incubation of semen with 5.0 microg/mL GA for 15 min completely stopped sperm motility. To test the reversibility of the GA effect on sperm motility, GA was removed after 30 min incubation and was replaced with fresh extender alone or with extender plus 5 mM caffeine, then incubated for another 15, 30, 45 or 60 min. The results showed that simply removing GA did not reverse the inhibitory effect on sperm motility, while adding caffeine partially reversed this inhibitory effect. However, the effect of 2.5 or 5.0 microg/mL GA was not reversed by caffeine. Considering the specificity of GA targeting to HSP90, the above observations suggested that HSP90 may play a crucial role in regulating porcine sperm motility.     

Peripheral plasma testosterone concentration and sexual behavior in young prenatally stressed boars

The purpose of this study was to examine the effects of stress induced physiological changes in the gestating sow on postnatal sexual and endocrine development of male offspring. Ten boars, ranging from 160 to 185 days of age, were randomly chosen from sows which had been maintained under either stress or control conditions during mid-gestation. Blood samples were collected weekly from each boar (minimum of four weeks) at 30 min intervals over a common six-hour period via an indwelling anterior vena cava cannula. Plasma testosterone concentrations were determined by radioimmunoassay. In order to ascertain degree of sexual behavior, boars were exposed weekly to gilts in estrus and a subjective score assigned. No differences (P>.10) were found between prenatally stressed and control boars in overall mean testosterone concentration or libido score. A significant (P<.05) decrease in plasma testosterone concentration was detected in boars over age. Results suggest that mid-gestational stress of gestating sows does not affect the testosterone concentration or sexual behavior of boar off-spring.     

Seasonality of LH,testosterone and sperm parameters in spider monkey males (Ateles geoffroyi)

There are no reported data on hormonal fluctuations in black‐handed spider monkey males. On previous research about the reproductive physiology of this monkey we have found that during the dry season females show ovulatory estrogen peaks and males present the best quality semen. As part of an ongoing research, in this study we assessed seasonal variations in the concentration of serum luteinizing hormone (LH) and testosterone (T) in three adult spider monkey males to corroborate the seasonal reproductive synchrony. At the same time sperm count and motility were evaluated to search for any correlation between those sperm parameters and hormonal concentrations. We took blood and semen samples (by electroejaculation) of anesthetized males throughout the rainy (June–September) and dry (October–May) months. Our results revealed that T and LH were higher throughout the dry season and there was a significant correlation between T concentration and sperm count. Although higher during the dry season, sperm motility tended to correlate with testosterone and LH levels. These results demonstrated that black‐handed spider monkeys have a tendency to show a seasonal pattern of reproduction being the dry season the most likely time to achieve fertilization. Am. J. Primatol. 71:427–431, 2009. © 2009 Wiley‐Liss, Inc.     

Immunological castration temporarily reduces testis size and function without long-term effects on libido and sperm quality in boars

The objective was to determine the effects of immunization against gonadotropin-releasing hormone on reproductive characteristics in boars. A total of 72 boars were used in a randomized design with three treatments: single immunization (SI) (10 weeks of age) or double immunization (DI) (10 and 15 weeks of age) with Improvest® and intact controls (no Improvest®; CNT) (n=24/group). At 10, 15, 20, 25 and 40 weeks of age, blood was collected and serum harvested to evaluate testosterone concentrations. Testosterone concentrations were less for DI boars compared with CNT boars and SI boars at 20 and 25 weeks (P<0.001), but not at 40 weeks of age. At week 25, 18 pigs (n=6/group) were sacrificed and testes were removed, weighed and measured, and seminiferous tubules were examined and scored using histological slides of testes parenchyma. A sample of neck fat was assessed for boar taint aroma. All testicular measurements and weights and seminiferous tubule scores were less for DI boars compared with SI and CNT boars (P<0.001). More (P<0.05) SI and CNT boars had detectable boar taint aroma than DI boars. Libido was assessed at 32, 36, 47, 60 and 63 weeks of age and semen collected at 60 weeks of age was analyzed for indicators of quality. There were no effects of treatment (P=0.41) or treatment by week (P=0.71) on libido. Semen volume, gel weight and total number of sperm cells, determined in a subset of boars (n=3/treatment), were not different among treatments. Sperm concentration was greater for DI than SI (P=0.01), and tended to be greater for DI compared with CNT (P=0.10). Sperm motility tended to be greater for DI boars compared with CNT boars (P=0.066). In conclusion, our results show that there are no long-term effects of immunocastration on reproductive characteristics in boars.     

Liquid storage of miniature boar semen.

The effects of liquid storage at 15 degrees C on the fertilizing ability of miniature pig semen were investigated. Characterization of ejaculated semen from 3 miniature boars was carried out. Semen volume and pH were similar among these boars. In one of the boars, sperm motility was slightly low, and sperm concentration and total number of sperm were significantly lower than in the others (P < 0.01). Seminal plasma of the semen was substituted with various extenders (Kiev, Androhep, BTS and Modena) by centrifugation and semen was stored for 7 days at 15 degrees C. Sperm motility was estimated daily at 37 degrees C. For complete substitution of seminal plasma, Modena was significantly more efficient than the other extenders (P < 0.001) in retaining sperm motility. Semen from each of the 3 miniature boars that had been stored for 5 to 7 days at 15 degrees C in Modena was used for artificial insemination of 15 miniature sows. The farrowing rates were 100, 100 and 60%, and litter sizes were 6.4 +/- 1.5, 5.8 +/- 0.8 and 5.0 +/- 1.0 for each boar semen, respectively. The boar that sired the smallest farrowing rate was the same one that showed lower seminal quality with respect to sperm motility, sperm concentration and total number of sperm. These results suggest that miniature boar semen can be stored for at least 5 days at 15 degrees C by the substitution of seminal plasma with Modena extender.     

Zona-penetration in vitro test for evaluating boar sperm fertility

We investigated the capacity of porcine sperm-zona binding and penetration by using bioassay to differentiate between spermatozoa from fertile and subfertile boars. Semen was collected from Large White boars grouped into categories of fertile and subfertile (n=5 per each group) according to the results of artificial insemination. Boars in both groups showed similarly hyperactivated sperm motility at insemination (44.72 and 43.03% respectively) regardless of the lower percentage of progressive motility observed in the ejaculates of subfertile boars. At in vitro insemination, a high proportion of the sperm population (43.76%) in the subfertile boars was without acrosomes, while in the fertile boars this proportion was only 24.35%. The sperm penetration rate of fertile boars reached 66.03% while that of subfertile boars was only 25.08%. In conclusion, the results of our study showed that the penetration rate by boar spermatozoa of the zona pellucida can be used to predict fertility and/or as an in vitro standard for describing porcine semen characteristics.     

Effects of centrifugation through three different discontinuous Percoll gradients on boar sperm function

In this study, different combinations of 2-step, discontinuous gradient centrifugation were used, consisting of three different combinations of isotonic Percoll (45/60, 60/75 and 45/90%) that allowed us to select different sperm subpopulations from fertile and normozoospermic boars. Our objective in this study is to evaluate the effects of centrifugation through three different discontinuous Percoll gradients on sperm function parameters (motility, viability, morphology, acrosome status, chromatin condensation, DNA fragmentation, ROS generation, tyrosine phosphorylation and intracellular calcium concentration) and the sperm penetrating capacity in an IVF system. All the Percoll treatments evaluated increased the percentage of spermatozoa with normal morphology, the proportion of un-damaged DNA, normal chromatin condensation, motion parameters measured by CASA and the percentage of capacitated spermatozoa with tyrosine phosphorylated proteins compared to control group. Finally, the in vitro oocyte penetrating capacity of boar spermatozoa was significantly affected by Percoll centrifugation. All the Percoll treatments increased the penetration rates and mean number of sperm per penetrated oocyte. Despite the efficiency of all three of the sperm treatments tested in selecting spermatozoa with improved sperm parameters and capacity to penetrate oocytes in vitro, the optimum performance of this system was demonstrated after preselecting spermatozoa by centrifugation on a discontinuous 45/90 Percoll gradient. The P45/90 treatment leads to obtain a higher percentage of spermatozoa which develop properly the capacitation process as it was shown measuring tyrosine phosphorylation and intracellular calcium concentration.     

Feeding with supplemental squalene enhances the productive performance in boars

In the present study, we investigated whether squalene could enhance the reproductive performance of boars. Boars at 12 months of age (weighting at 130+/-5 kg) were randomly allocated into four groups and treated with basal diet (control) or with basal diet supplemental squalene at 10, 20 or 40 mg squalene/kg/day for 60 days. The impact of squalene feeding on the reproductive performance and the levels of serum leptin and testosterone were evaluated before and after feeding. Feeding with squalene at 10 mg/kg/day neither significantly increase the reproductive performance and serum testosterone levels, nor reduced the levels of leptin in boars. Surprisingly, feeding with supplemental squalene at 20 or 40 mg/kg/day significantly improved the reproductive performance as evidenced by dramatically reduced the time for mating, increased semens' volume and motility, and increased the size of litter as compared with that in controls. Furthermore, feeding with a higher dose of squalene significantly reduced the levels of serum leptin, accompanied by elevated levels of testosterone, as compared with that in controls. These data clearly indicate that feeding with squalene can improve the reproductive performance in boars. Therefore, dietary supplementation with squalene may increase the productivity of pig industry and potentially benefit for other animal production.