Vitamin E and organic selenium enhances the antioxidative status and quality of chicken semen under high ambient temperature

1. The effects of supplemental dietary vitamin E and organic selenium (Se), and their combination, on improving semen quality characteristics and antioxidative status were investigated in cockerels exposed to high ambient temperature.

2. A total of 36 Egyptian local cross males, 40 weeks old, were housed individually in cages in an open-sided building (average daily temperature ranged from 33 to 36°C and relative humidity from 60 to 70%). Birds were divided randomly into 4 experimental treatments (n?=?9 each): (1) control (basal diet without any supplementation with vitamin E or Se); (2) vitamin E (basal diet +200?mg α-tocopherol acetate/kg diet); (3) Se (basal diet +0·3?mg organic Se/kg diet); and (4) vitamin E?+?Se (basal diet +200?mg α-tocopherol acetate/kg diet +0·3?mg organic Se/kg diet).

3. Under heat stress conditions, inclusion of vitamin E and/or organic Se in the diets enhanced the semen quality traits, including the spermatozoa count and motility, and reduced the percentage of dead spermatozoa.

4. A combination of 200?mg/kg vitamin E with 0·3?mg/kg organic Se reduced the thiobarbituric acid reactive substance concentration in seminal plasma samples to about 28% of the controls; and also enhanced the seminal plasma glutathione peroxidase activity by two-fold compared with controls.

5. It was concluded that dietary vitamin E in combination with organic Se has a synergistic effect in minimising lipid peroxidation and improving the antioxidative status in seminal plasma of the domestic fowl, which probably translated into enhanced spermatozoa count, motility and reduced percentage of dead spermatozoa under heat stress conditions.     

Effects of supplemental vitamin E during the mature period on the reproduction performance of Taiwan Native Chicken cockerels

One-day-old Taiwan native male chicks were fed with maize-soybean rearing diets without supplemental vitamin E to 23 weeks of age. From 23 to 52 weeks of age, the cockerels (n = 90) were assigned at random to 5 dietary treatments and fed with maize-soybean diets supplemented with 0, 20, 40, 80 and 160 mg/kg of vitamin E (dl-alpha-tocopherol acetate). Pullets (225) of the same age were fed with standard diets throughout. They were artificially inseminated with one dose of 0.04 ml/bird intact and 5-fold diluted pooled semen at 31 to 43 weeks of age and at 49 weeks of age, respectively. The criteria evaluated included: semen quality, fertility and maximum and effective duration of fertility, blood characteristics, body and testes weight. Supplemental vitamin E did not affect cockerels' effective duration of fertility and percentage of fertility. However, when pullets were inseminated with diluted semen, supplementing 160 mg/kg vitamin E increased the maximum duration of fertility at 49 weeks of age. Cockerels receiving 40 to 160mg/kg supplements had higher sperm viability and motility after 39 weeks of age and those fed 80 mg/kg had higher sperm concentration at 39 weeks of age. Cockerels receiving supplements of more than 40 mg/kg vitamin E had higher body weight gain. Plasma cholesterol and testosterone were not affected by supplemental vitamin E. However, plasma luteinising hormone (LH) concentration was lower in cockerels fed 160 mg/kg. Lack of supplemental vitamin E over 39 weeks was associated with lower semen quality but did not reduce the proportion of fertile eggs laid by inseminated hens, perhaps because the insemination dose compensated for low sperm quality. We found that the maximum duration of fertility might be improved by supplementing 160 mg/kg vitamin E at 49 weeks of age.     

补饲硒和维生素E对高温季节山羊精液品质、抗氧化酶活性及热休克蛋白表达的影响

试验旨在研究在海南夏季高温饲养条件下,山羊补饲硒和维生素E对其精液品质、抗氧化酶活性及热休克蛋白表达的影响。选用16只健康状况良好、体重相近的海南黑山羊成年公羊,随机分成4组,分别饲喂基础日粮(对照组)、基础日粮+0.5mg/kg Se(Se组)、基础日粮+100mg/kg VE(VE组)和基础日粮+0.5mg/kg Se+100mg/kg VE(Se+VE组),试验期93 d。试验期结束前1周采集精液,评价精液品质、测定精浆抗氧化酶活性和精液热休克蛋白表达。结果表明,与对照组相比,补饲Se和VE对海南黑山羊射精量影响差异不显著(P>0.05);补饲Se和VE能显著提高海南黑山羊的精子密度和精子活力(P<0.05),极显著降低精子畸形率(P<0.01);极显著增加精浆谷胱甘肽过氧化物酶活性和总抗氧化能力(P<0.01),显著增加超氧化物歧化酶和过氧化氢酶活性(P<0.05),显著降低丙二醛浓度(P<0.05);补饲Se和VE还极显著降低了精液HSP70和HSP90mRNA的表达水平(P<0.01),但各补饲组之间对精液品质改善效果和热休克蛋白表达丰度的影响存在一定差异。综上所述,补饲Se和VE有助于提高热带地区夏季高温季节山羊精子密度、精子活力,降低畸形率,同时还能增强精浆中抗氧化酶的活性,改善精液品质,进而起到缓解环境热应激的效果。     

Effect of Se and VE Supplement on Semen Quality,Antioxidant Enzyme Activities and Heat Shock Protein Expression of Goat in High Temperature Season

This experiment was conducted to study the effect of selenium and vitamin E supplement on semen quality, antioxidant enzyme activities and heat shock protein expression of goat in Hainan high temperature season.16 adult Hainan Black goat with good health and approximate weight were randomly divided into 4 groups, fed with basal diet(control group), basal diet+0.5 mg/kg Se(Se group), basal diet+100 mg/kg VE(VE group), and basal diet+0.5 mg/kg Se+100 mg/kg VE(Se+VE group), respectively.The experimental period was 93 d.Semen samples were collected in the last week of the experiment on two consecutive days.The semen quality, antioxidant enzyme activities and heat shock protein expression were analyzed.The results showed that compared with control group, the ejaculate volume was not significantly affected by Se or VE supplement(P>0.05).Sperm density and sperm motility were increased significantly by Se and VE supplement(P<0.05), and the abnormal rate was decreased extremely significantly(P<0.01).The goats fed with Se and VE also had higher activities of GSH-Px(P<0.01), SOD(P<0.05), CAT(P<0.05) and T-AOC(P<0.01), and lower MDA concentration in seminal plasma(P<0.05).The relative expression levels of HSP70 and HSP90 mRNA in supplement groups were decreased extremely significantly(P<0.01).However, there were some certain differences between the Se and VE supplement groups on semen quality and heat shock protein expression.In conclusion, the supplementation of Se and VE could help to improve goat semen quality by increasing the sperm density, sperm motility, the antioxidant enzyme activities, and decreasing the abnormal rate in hot season of Hainan.Finally, Se and VE supplement had good effects on relieving the environment heat stress.     

Effect of vitamins,probiotics and protein level on semen traits and seminal plasma biochemical parameters of post-moult male broiler breeders

1. A study was designed to investigate the comparative effect of supplementary vitamin E, vitamin C, probiotics and dietary crude protein concentration on semen traits and seminal plasma biochemical parameters in male broiler breeders after Zn-induced moulting.

2. A total of 180 male broiler breeders were induced to moult at 65 weeks of age by mixing ZnO in diet at the rate 3000?mg/kg of feed. After moulting, the males were divided into six groups that were given feed supplemented with: vitamin C (500?IU/kg), vitamin E (100?IU/kg), protein (140?g CP/kg), probiotics (50?mg/kg) and the combination of these components, while one group was kept as a control. Semen samples were collected weekly and semen volume, spermatozoa concentration, motility and dead spermatozoa percentage were determined. Seminal plasma was separated to determine the concentration of total antioxidant capacity (TAC), homocysteine, paraoxonase (PON1), arylesterase, ceruloplasmin, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities.

3. Semen volume was significantly higher in the vitamin E and C groups compared to the control. Spermatozoa motility was higher in the vitamin E group and dead spermatozoa percentage was significantly lower in the vitamin C group compared with the control group.

4. Seminal plasma TAC was higher in the vitamin E group, homocysteine was lower in the vitamin C and E groups. PON1 was higher in the combination group. Arylesterase increased significantly in the vitamin C and combination groups over time. Seminal plasma AST was significantly lower in the vitamin C and E supplemented groups whereas ALT decreased significantly only in the vitamin E group compared with the control. Higher concentrations of ceruloplasmin were observed in the combination group compared with the other treatments.

5. It was concluded that additional vitamin E and C or their combination was the most potent nutrient treatment for improving the semen traits and seminal plasma biochemical characteristics in male broiler breeders after Zn-induced moulting.     

影响意大利水牛精液品质的相关因素分析

本研究旨在通过对意大利水牛精液品质分析、睾丸周径测量、精浆中的氧化应激水平检测和精子活力相关基因表达情况来探究影响精液质量的相关因素。试验检测了6头意大利水牛精液的活力、畸形率、采精量,并在测量其阴囊周径后进行相关性分析;检测了意大利水牛精浆中氧化应激水平指标（丙二醛（MDA）、总超氧化物歧化酶（T-SOD）、谷胱甘肽过氧化物酶（GSH-Px））并进行了相关性分析;分析了意大利水牛精子β-微管蛋白-2c（TUBB2C）、外周致密纤维2（ODF2）、筑丝蛋白2（TEKT2）、筑丝蛋白4（TEKT4）基因的表达定量及相关性。结果显示,意大利水牛阴囊周径与精液产量、活力、畸形率之间相关系数分别为0.423（P>0.05）、0.750（P<0.01）、-0.827（P<0.01）,即阴囊周径与精子活力呈显著正相关关系、与畸形率呈显著负相关关系;意大利水牛精子活力与MDA、T-SOD、GSH-Px指标之间相关系数分别为-0.522（P<0.05）、0.333（P>0.05）、0.474（P<0.05）,即精子活力与MDA含量之间存在显著负相关关系,与GSH-Px活性之间存在显著正相关关系,与T-SOD活性相关性不显著;意大利水牛精子活力与TUBB2C、ODF2、TEKT2、TEKT4基因指标之间相关系数分别为0.930（P<0.01）、0.726（P<0.01）、0.924（P<0.01）、0.839（P<0.01）,即精子活力与以上基因表达量存在显著正相关关系。本研究结果为了解影响意大利水牛精液质量的因素提供参考,也为意大利种公水牛的筛选提供理论依据。     

Analysis of Related Factors in Semen Quality of Italian Buffaloes

This study was aimed to investigate the factors affecting semen quality by analyzing the semen quality of Italian buffaloes,measuring the circumference of the testis,the level of oxidative stress in seminal plasma and the expression of genes related to sperm motility.In this study,the sperm vitality,deformity rate and semen collection of 6 Italian buffaloes were tested,and the correlation analysis was carried out after measuring scrotal circumference.The oxidative stress level of seminal plasma in Italian buffaloes was tested,including malondialdehyde (MDA),total superoxide dismutase (T-SOD) and glutathione peroxidation (GSH-Px).The expression of β-tublin-2c (TUBB2C),outer dense fibres 2 (ODF2),tektin-2 (TEKT2),tektin-4 (TEKT4) genes and their correlation with sperm in Italian buffaloes were analyzed.The results showed that the correlation between scrotal circumference and semen yield,vigor and deformity rate was found,the coefficients were 0.423 (P>0.05),0.750 (P<0.01) and -0.827 (P<0.01),which showed a significant positive correlation between scrotal circumference and sperm motility,and a significant negative correlation with the deformity rate.The oxidative stress level in seminal plasma of Italian buffaloes was tested,the correlation coefficients of MDA,T-SOD and GSH-Px with sperm motility were -0.522 (P<0.05),0.333 (P>0.05) and 0.474 (P<0.05),respectively.There was a significant negative correlation between sperm motility and MDA content,a significant positive correlation between GSH-Px activity and sperm motility,no significant correlation with T-SOD activity.The correlation coefficients between sperm motility and TUBB2C,ODF2,TEKT2 and TEKT4 genes were 0.930 (P<0.01),0.726 (P<0.01),0.924(P<0.01) and 0.839 (P<0.01),respectively.There was a significant positive correlation between sperm motility and the expression of four genes.The results provided a reference for understanding the factors affecting the semen quality of Italian buffaloes,and also provided a theoretical basis for the selection of Italian bull.     

In vivo adverse effects of alpha‐tocopherol on the semen quality of male bucks

Oxidative stress has detrimental effects on semen quality during spermatogenesis and semen processing for artificial insemination. This work was conducted to study the effect of different levels of vitamin E on the semen traits, oxidative status and trace minerals in Beetal bucks. Thirty‐six bucks of similar body weight and age (1 year) were randomly divided into four groups. One group was kept as control with no supplementation (group 1), and the others were supplemented with 200 (group 2), 400 (group 3) and 800 IU (group 4) vitamin E/animal/day for 2 months. At the end of the experiment, semen samples were collected and evaluated. Seminal plasma was separated to study the concentration of superoxide dismutase (SOD), glutathione peroxidase (GPx), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and trace minerals (Zn, Cu, Mn and Fe). Group 3 showed significantly higher (p < 0.05) semen volume and per cent motility and lower dead sperm percentage compared to control group. Superoxide dismutase, GPx, Zn, Cu and Mn were higher in the same group. The level of AST decreased in group 3 without any change on the concentration of ALT. It is suggested that vitamin E at the rate of 400 IU/buck/day supported higher semen volume, per cent motility, per cent live spermatozoa, antioxidants (SOD, GPx) and trace mineral levels (Zn, Cu, Mn) in the seminal plasma. The increased supplementation from 0 to 400 showed a general increasing trend in improving semen quality. However, the dose of 800 IU/kg had no useful effect in further improving the semen quality.     

维生素E对秦川牛冷冻精液的抗氧化保护作用

本试验旨在研究维生素E对秦川牛细管冷冻精液品质和精浆中抗氧化酶活性的影响.在稀释液中分别添加0.00、0.02、0.04、0.06、0.08、0.10 mg·mL~(-1)的维生素E,将细管冻精解冻(37.5℃,30 s)后用伟力精子分析系统分析精子活力和精子顶体完整率等标准参数,并用相关试剂盒测定精浆中的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)、谷胱甘肽还原酶(GR)和过氧化氢酶(CAT)的活性.结果表明:当维生素E添加量为0.04mg·mL~(-1)时,精子活力与对照组相比差异显著(P<0.05),SOD、GSH、CAT 3种酶活性与对照组相比差异显著(P<0.05).当维生素E的添加量为0.06 mg·mL~(-1)时,冷冻-解冻后精子活力显著高于对照组(P<0.05),顶体完整率极显著高于对照组(P<0.01);GR和CAT酶活性与对照组相比均差异显著(P<0.05).当维生素E的添加量为0.08 mg·mL~(-1)时,精子活力、活率及顶体完整率与对照组相比差异显著(P<0.05),GSH与对照相组比差异极显著(P<0.01),GR及CAT与对照组相比差异显著(P<0.05).在冷冻精液稀释液中添加维生素E可以有效提高冷冻-解冻后秦川牛精液品质及精浆中SOD、CAT、GSH、GR酶的活性,稀释液中维生素E的适宜添加量为0.04～0.08 mg·mL~(-1).     

Effect of supplementation with the reduced form of coenzyme Q10 on semen quality and antioxidant status in dogs with poor semen quality: Three case studies

Oxidative stress owing to an imbalance between reactive oxygen species and antioxidants, such as coenzyme Q10 (CoQ10), is a major contributor to male infertility. We investigated the effects of the reduced form of CoQ10 (ubiquinol) supplementation on semen quality in dogs with poor semen quality. Three dogs received 100 mg of ubiquinol orally once daily for 12 weeks. Semen quality, serum testosterone, and seminal plasma superoxide dismutase (SOD) activity were examined at 2-week intervals from 2 weeks before ubiquinol supplementation to 4 weeks after the treatment. Ubiquinol improved sperm motility, reduced morphologically abnormal sperm, and increased seminal plasma SOD activity; however, it had no effect on testosterone level, semen volume, and sperm number. Ubiquinol supplementation could be used as a non-endocrine therapy for infertile dogs.     

Characterization of semen collected by pharmacologically induced ejaculation from a stallion with seminal vesiculitis

The present study compared the quality of sperm collected by artificial vagina or pharmacologically induced ejaculation from a 10-year-old thoroughbred stallion with seminal vesiculitis. The pharmacological protocol involved intravenous administration of detomidine (0.01 mg/kg) and oxytocin (20 IU) and successfully induced ejaculation in all attempts of semen collection. Sperm motility, plasma membrane and acrosome integrity (PMAI), reactive oxygen species (ROS) levels, polymorphonuclear neutrophil (PMN) percentage, and bacterial profiles of fresh and cooled semen (5°C for 24 hr) were evaluated. Semen obtained by the pharmacological method presented reduced seminal volume, decreased PMN percentage and superior sperm motility in cooled samples. Moreover, higher PMAI and lower ROS levels were observed in semen collected by the pharmacological method. Therefore, pharmacologically induced ejaculation is an alternative to obtain semen with minimal contamination and with sperm of superior quality and longevity from stallions with seminal vesiculitis.     

Effect of dietary selenium and vitamin E on the ultrastructure and ATP concentration of boar spermatozoa, and the efficacy of added sodium selenite in extended semen on sperm motility

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.     

Influence of vitamin E and vitamin E-selenium combination on arginase activity,nitric oxide level and some spermatological properties in ram semen

The effects of vitamin E and vitamin E-selenium combination on seminal plasma arginase activity and nitric oxide level and some spermatological properties in rams were investigated in this study. For control group, animals were injected intramuscularly with physiological saline. For vitamin E group, rams were injected intramuscularly with 300 mg/ram vitamin E. For vitamin E + selenium group, animals were injected intramuscularly with 5 ml/ram vitamin E + selenium. The semen was collected by artificial vagina at 1st, 4th, 24th, 48th and 72nd hr after administration in each group. Significant decreases in seminal plasma arginase activity (at 1st, 24th and 48th hr), nitric oxide level (at 72nd hr) and abnormal sperm rate (at 1st, 24th and 72nd hr), and significant increases in semen volume (at 24th hr), semen mass activity (at 24th and 48th hr), sperm motility (at 24th, 48th and 72nd hr) and concentration (at 1st hr) were observed in vitamin E group compared with control group. Similarly, significant increase in semen volume (at 1st, 24th and 48th hr), mass activity, (at 48th hr), motility (at 48th and 72nd hr) and concentration (at 4th, 24th and 48th hr), and significant decrements in abnormal sperm rate (at 1st, 24th, 48th and 72nd hr), seminal plasma nitric oxide level (at 1st, 4th, 24th and 48th hr) and semen pH (at 24th and 48th hr) were detected in vitamin E + selenium group in comparison to the control group. As a result, it is suggested that vitamin E and/or vitamin E + selenium applications may improve reproductive performance.     

长白公猪血清和精浆中元素含量对精液品质的影响

本试验旨在研究不同精液品质特征长白公猪血清和精浆中元素含量的差异,分析血清和精浆中元素含量对精液品质的影响。基于107头长白公猪的1402次精液品质记录,将公猪群按照精液可利用率划分为3组:低利用率组(利用率<60%,n=21)、中等利用率组(60%≤利用率≤80%,n=27)和高利用率组(利用率>80%,n=59)。采集每头长白公猪的血清和精浆样品,采用电感耦合等离子体质谱法检测血清和精浆中营养元素和毒性元素含量。结果表明:1)低利用率组长白公猪的有效精子数和精子活力显著低于中等利用率和高利用率组(P<0.01),精子畸形率显著高于中等利用率和高利用率组(P<0.01)。2)不同利用率组间长白公猪的血清和精浆元素含量无显著差异(P>0.05);但在血清和精浆元素含量与精液品质参数相关性分析中发现,精浆铅元素含量与精子活力呈显著负相关(P<0.05),与精子畸形率呈显著正相关(P<0.05)。3)对精浆铅含量分组进一步分析发现,精浆铅含量为0μg/L时长白公猪的精子畸形率显著低于精浆铅含量>10.0μg/L时(P<0.05),精子畸形率降低约6.11%。总的来说,长白公猪精浆中毒性元素铅的存在会通过损害精子活力和形态,影响公猪精液品质。     

Total antioxidant capacity and protein peroxidation intensity in seminal plasma of infertile and fertile dogs

The aim of this study was to evaluate the total antioxidant capacity and protein peroxidation intensity in seminal plasma of infertile and fertile dogs. The study was conducted on 10 infertile and 10 fertile dogs of various breeds. Infertility was defined as conception failure at least three matings with different bitches. Semen was collected by manual manipulation. The sperm concentration and motility parameters were evaluated using CASA Hamilton Thorne, Vers. IVOS 12.3. The morphology of spermatozoa and the percentage of live and dead sperm cells were assessed microscopically, total antioxidant capacity and the content of SH‐groups in seminal plasma were determined spectrophotometrically, the contents of protein peroxidation markers in seminal plasma, bityrosine and formylokinurenine, were determined using spectrofluorimetric methods. Sperm concentration and total sperm count were significantly (p < 0.05) lower in infertile dogs than in fertile dogs (99.92 ± 3 0.05 × 10⁶/ml vs. 282.07 ± 48.27 × 10⁶/ml; 214.19 ± 114.74 × 10⁶ vs. 747.57 ± 210.94 × 10⁶, respectively). The percentage of spermatozoa with normal morphology and the most determined motility parameters differed significantly (p < 0.05) between both groups. The mean values of total antioxidant capacity in the seminal plasma were significantly (p < 0.05) lower (19.95 ± 20.94 vs. 25.66 ± 23.18 µmol/g protein), whereas the mean contents of bityrosine and formylokinurenine in seminal plasma were significantly (p < 0.05) higher in infertile dogs than in fertile dogs (3.71 ± 4.83 µg/mg protein vs. 1.55 ± 2.00 µg/mg protein and 0.37 ± 0.45 µg/mg protein vs. 0.14 ± 0.08 µg/mg protein, respectively). In conclusion, the obtained results suggest that the poor semen quality and infertility in dogs could be associated with lowered total antioxidant capacity and increased protein peroxidation in seminal plasma as a consequence of oxidative stress.     

A comparison of electroejaculation and epididymal sperm collection techniques in stallions

The purpose of this study was to evaluate 2 methods of semen collection that could be used as terminal procedures in stallions with irreparable conditions, such as fractures or colic. Electroejaculation was attempted under general anesthesia. Forty-eight hours later, the ponies were castrated and 2 different epididymal sperm collection techniques were attempted by using a flushing or floating method. Additionally, the effect of supplemental seminal plasma was evaluated. Experimentally, electroejaculation was found to be a safe but ineffective method of terminal semen collection. Viable sperm cells were successfully recovered with both types of epididymal collection. The flotation method was least cumbersome and showed a tendency to be superior to flushing in terms of sperm motility and percentage of cells passing through glass wool/sephadex filtration, although differences did not reach significance. The addition of seminal plasma to epididymal spermatozoa prior to cryopreservation was of no value. In conclusion, either method of epididymal sperm collection is an acceptable method of terminal semen collection.     

Organic zinc and copper supplementation on antioxidant protective mechanism and their correlation with sperm functional characteristics in goats

Trace minerals feeding had significant effects on sperm production and fertility with better absorption and proper utilization within the body for optimum reproductive function. Several studies have shown that more influenced trace elements in the diets of animals are copper (Cu) and zinc (Zn). Bucks showing deficiency of this mineral might affect the quality of semen production which in turn would affect the fertility. This experiment was thus designed to test the effects of organic Cu and Zn supplementation on antioxidants enzyme activities and sperm functional attributes in fresh semen of bucks. Forty bucks (n = 40, Aged 5 months) were assigned to ten groups of four animals in each group, supplemented (for a period of 8 months) with different levels of organic Zn: 20 mg (T2), 40 mg (T3) and 60 mg (T4), organic Cu: 12.5 mg (T5), 25 mg (T6), 37.5 mg (T7) and combined organic Zn and Cu: 20 + 12.5 mg (T8), 40 + 25 mg (T9), 60 + 37.5 mg (T10), respectively, per kg dry matter and no additional mineral diet (control; T1). One hundred and sixty semen samples were collected through electro‐ejaculator and analysed for sperm quantity, quality, acrosome intactness and plasma membrane integrity and correlated with the catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase enzyme activities in seminal plasma. The results indicated organic Cu and zinc supplemented bucks produced more sperm cells, had higher sperm concentrations, maintained higher (p < .01) sperm livability, plasma membrane and acrosome integrities, more motility and velocity. The increased antioxidant enzyme activities, reduced oxidative stress and lowered lipid peroxidation were positively correlated (p < .05) with the sperm functional attributes. In conclusion, organic Cu and Zn supplement to male goats showed protective roles against oxidative damage and maintained better fresh semen characteristics.     

奶牛性控精液与常规精液抗氧化物酶活性比较分析

通过比较常规精液和性控精液在活力、顶体完整率及精浆中四种抗氧化酶类活力的差异,对性控精液的品质进行综合评定。对用流式细胞仪分离后的性控精液和常规精液细管解冻(37.5℃,30s),分别分析精子活力和精子顶体完整率,并测定精浆中的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)、谷胱甘肽还原酶(GR)和过氧化氢酶(CAT)的活性。结果表明性控精液精子在活力显著低于常规精液,且精浆中四种抗氧化酶类的活力也都显著低于常规精液。     

不同季节肉用种公牛精液品质与生理常值、血清及精清生化指标相关关系研究

以利木赞和夏洛来种公牛为试验牛，研究不同季节对其精液品质、冻精产量、生理常值、血清及精清生化指标的影响。结果表明：（1）与其它季节相比，夏季肉用种公牛的原精活力、冻精活力、活精于百分数、顶体完整率均显著降低，而精于畸形率显著升高（P〈0．05）。精子密度以秋季最低。（2）冻精产量夏季处于最低水平，平均每月仅为7388支。（3）夏季肉用种公牛的血清LH、睾酮、T3、皮质醇的含量分别为48．12、4．15、1．11、4．89ng／L，显著低于春、秋、冬季（P〈0．01）。（4）夏季肉用种公牛血清钠、钾、钙、镁均处于最低水平，显著低于其它季节（P〈0．01）。（5）夏季肉用种公牛精清睾酮水平明显下降，显著低于春季、秋季和冬季。（6）精清钠、钾、钙夏季含量最低，分别为83．12mmol／L、18，77mmol／L和4．63mmol／L；精清镁含量由春季至秋季呈显著下降趋势；精清磷含量夏季最高，为4．65mmol／L，显著高于春季、秋季和冬季。（7）肉用种公牛精清ALP夏季含量为1784．3U／L，显著高于春、秋两季，但低于冬季。（8）精清睾酮与精液品质各指标存在显著相关；血清生化指标与精清生化指标存在显著相关；直肠温度、呼吸频率与精液品质及血清生化指标存在显著相关性，与精清生化指标相关性不大。     

Removal of Seminal Plasma Enhances Membrane Stability on Fresh and Cooled Stallion Spermatozoa

Fertility is reduced after semen cooling for a considerable number of stallions. The main hypotheses include alterations in plasma membrane following cooling and deleterious influence of seminal plasma. However, interindividual variability is controversial. We hypothesized that the removal of seminal plasma could enhance motility in some ‘poor cooler’ stallions, but could also affect, negatively or positively, membrane quality in some stallions. This study examined the effect of centrifugation, followed or not by removal of seminal plasma, on parameters indicating semen quality after 48 h at 4°C: motility, plasma membrane integrity as evaluated by hypo‐osmotic swelling test, acrosome integrity and response to a pharmacological induction of acrosome reaction using ionophore A23187. Sixty‐six ejaculates from 14 stallions were used, including stallions showing high or low sperm motility after cooled storage. Centrifugation without removal of seminal plasma did not affect sperm parameters. Removal of seminal plasma did not affect motility, but significantly stabilized sperm membranes, as demonstrated by a higher response to the osmotic challenge, and a reduced reactivity of the acrosome. Moreover, for the same semen sample, the response to an induction of acrosome reaction was significantly higher when the induction was performed in the presence of seminal plasma, compared with the induction in the absence of seminal plasma. This was observed both for fresh and cooled semen. When the induction of acrosome reaction with ionophore A23187 is used to evaluate sperm quality, care must therefore be taken to standardize the proportion of seminal plasma between samples. For the 10 stallions serving at least 25 mares, the only variable significantly correlated with fertility was motility. The influence of membrane stabilization regarding fertility requires further investigations.