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.     

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).     

Vitamin E supplementation reduces dexamethasone-induced oxidative stress in chicken semen

1. We examined the effects of supplemental dietary vitamin E (Vit E) on semen quality and antioxidative status in male domestic fowls exposed to oxidative stress induced by synthetic glucocorticoid, dexamethasone (DEX) injection. 2. Thirty-six Egyptian local cross males, 42 weeks old, were housed individually in cages in an open-sided building under 16 h light:8 h dark and were provided with commercial feed and water ad libitum. Birds were divided into 4 groups: DEX (4 mg/bird/d), Vit E (200 mg/kg diet), DEX + Vit E (4 mg/bird/d + 200 mg/kg diet, respectively) and control, n = 9. All treatments lasted for 7 continuous days. 3. Oxidative stress induced by injection of DEX (4 mg/bird/d) resulted in decreased sperm count and motility correlated with an increased percentage of dead sperms. Vit E (200 mg/kg diet) enhanced sperm count and viability when supplemented to stress-induced birds, compared to DEX treatment alone. 4. In seminal plasma, low calcium concentration, high lipid peroxidation and reduced activity of glutathione peroxidase were associated with the oxidative stress. Vit E reduced lipid peroxidation in the seminal plasma. 5. In conclusion, excessive supplemental dietary Vit E improved semen quality when cockerels were subjected to stress conditions. It increased both sperm count and motility, reduced the percentage of dead sperm and enhanced the antioxidative status of seminal plasma.     

Efficacy of Low‐Dose Human Chorionic Gonadotropin Therapy in Dogs with Spermatogenic Dysfunction: A Preliminary Study

Human chorionic gonadotropin (hCG) is a glycoprotein used in the treatment of spermatogenic dysfunction. However, previous studies performed in dogs show that repeated administration of large doses of hCG produces antibodies against hCG. In this study, we examined the efficacy of low‐dose injections of hCG in four male dogs with spermatogenic dysfunction and low plasma testosterone (T) levels. We administered 100 IU hCG per animal, five times at 3‐day intervals, and evaluated the changes in semen quality and plasma T levels. The total number of sperm in ejaculate, the percentage of progressively motile sperm and the plasma T levels had increased by 3–5 weeks after the first injection of hCG in three of the four dogs, but were unchanged in the fourth dog. These findings indicate that temporary improvement of the semen quality of dogs with spermatogenic dysfunction and low plasma T levels is possible after five low‐dose injections of hCG.     

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.     

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.     

Effect of Post‐Thaw Addition of Seminal Plasma on Motility,Viability and Chromatin Integrity of Cryopreserved Donkey Jack (Equus asinus) Spermatozoa

Pregnancy rates in donkeys after artificial insemination with cryopreserved semen are still low, compared to the horse species. Addition of autologous seminal plasma to frozen‐thawed semen appeared to improve pregnancy rates. The aims of this study were to evaluate (1) sperm motility and plasma membrane integrity after thawing (T0) and after one and 2 h (T1 and T2) of post‐thaw incubation in either 0% (SP0) or 70% (SP70) autologous seminal plasma and (2) sperm motility, plasma membrane integrity and DNA quality (%COMP‐αt) after thawing (T0) and after 2 and 4 h (T2 and T4) of post‐thaw incubation in either 0% (SP0), 5% (SP5) or 20% (SP20) homologous seminal plasma. In experiment 1, seminal plasma decreased total and progressive sperm motility and plasma membrane intact spermatozoa immediately after dilution and at all following time points (p < 0.05). In experiment 2, total and progressive motility did not differ between treatments immediately after dilution and between SP0 and SP5 at T2, while they were lower in both SP5 and SP20 than in SP0 at T4. Plasma membrane intact sperm cells did not differ between SP0 and SP5 and were lower in SP20 at all time points. DNA quality was not affected by treatment immediately after dilution and was significantly worse for SP20 after 4 h of incubation (p < 0.05). The post‐thaw addition of seminal plasma at the tested concentrations did not improve donkey frozen semen characteristics in vitro over time.     

Superoxide dismutase and catalase activities in the seminal plasma of normozoospermic and asthenozoospermic Beagles

We measured the blood plasma testosterone (T) levels and superoxide dismutase (SOD) and catalase activities in the seminal plasma of the ejaculates of 5 normal (2-5 years old) and 5 asthenozoospermic (AZ-) (3-5 years old) Beagles. Sperm ejaculated by AZ-dogs was incubated for 3 hr in Eagle's MEM only (controls) or Eagle's MEM containing 100 units/ml of SOD or catalase. Sperm motility was examined during incubation. The mean (+/- SE) plasma T level of the AZ-dogs (1.2 +/- 0.2 ng/ml) was significantly lower than in the normal dogs (2.5 +/- 0.2 ng/ml) (P<0.005). The mean (+/- SE) seminal plasma SOD and catalase activities (18.8 +/- 1.9 and 0.5 +/- 0.1 unit/g protein, respectively) were significantly lower in the AZ-dogs than in the normal dog (43.3 +/- 2.5 and 2.2 +/- 0.4 unit/g protein, respectively) (P<0.001 and 0.01, respectively). The motility of sperm incubated in Eagle's MEM containing SOD or catalase was significantly higher than that of control sperm incubated in only Eagle's MEM after 2 or 3 hr of incubation (P<0.05). The results of this study indicate that poor T secretion by the testes and low antioxidant enzyme activities are related to AZ in the dog.     

Effect of seminal plasma concentration and various extenders on postthaw motility and glass wool-Sephadex filtration of cryopreserved stallion semen

OBJECTIVE: To compare the effect of semen extender and seminal plasma on postthaw motility and filtration through a glass wool-Sephadex (GWS) filter for frozen stallion semen. SAMPLE POPULATION: 7 stallions from which we collected > or = 3 ejaculates/stallion. PROCEDURES: 4 experiments were conducted to evaluate postthaw quality of frozen stallion semen. Kenney extender was compared with glucose-EDTA extender by use of various dilution rates that resulted in differing concentrations of seminal plasma. Stallions known to produce semen with poor postthaw quality were used to investigate whether a particular extender or dilution rate could improve ability of such semen to survive freeze-thaw procedures. RESULTS: Use of Kenney extender as the centrifugation extender significantly improved postthaw motility and GWS filtration, compared with glucose-EDTA. Extending semen at a dilution of 1:3 was significantly better than 1:1 for both motility and GWS filtration. In addition, including seminal plasma at a concentration of 5% in the cryopreserved semen resulted in significantly higher yield of spermatozoa after GWS filtration, compared with complete removal of SP or use of seminal plasma at 25%. Lastly, semen with poor postthaw quality had significantly improved postthaw quality in regard to motility and GWS filtration when semen was frozen with seminal plasma at a concentration of 5%, compared with semen frozen with seminal plasma at a concentration of 25%. CONCLUSIONS AND CLINICAL RELEVANCE: Use of Kenney extender at a high dilution (> or = 1:3) immediately after collection of semen can improve postthaw quality of frozen stallion semen.     

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.     

Therapeutic effect of aromatase inhibitor in two azoospermic dogs with high plasma estradiol-17beta levels

Two azoospermic dogs with high plasma estradiol-17 beta (E(2)) levels were subcutaneously injected with an aromatase inhibitor (AI), 4-androstene-4-ol-3,17-dione, 2 mg every other day for 4 weeks. Before the AI treatment the plasma E(2) levels of the two dogs (21 and 22 pg/ml, respectively) were higher than those of 2 normal dogs (8.1 and 12.3 pg/ml), and they fell to 11-17 pg/ml between 1 and 4 weeks after the start of AI treatment. The plasma testosterone levels after the start of AI treatment had increased to 2.1-3.1 ng/ml. A small number of sperm were detected in the semen of the two dogs between 3 and 6 weeks after the start of AI treatment. These results indicate that the testicular function of infertile dogs with high plasma E(2) levels can be temporarily improved by AI therapy.     

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.     

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.     

Seasonal variation in testicular blood flow dynamics and their relation to systemic and testicular oxidant/antioxidant biomarkers and androgens in rams

The environmental temperature increased during summer and decreased during winter to the limits that might negatively affect animal and human reproduction. The responses of Egyptian rams to either hot or cold climatic conditions were studied in six mature rams subjected to weekly testicular Doppler ultrasonographic examination, blood sampling, seminal plasma collection and semen evaluation. The maximum environmental temperature and the relative humidity were used to classify the climatic condition according to the heat stress equation of sheep into hot months where temperature–humidity index (THI) was >26 (31.67 ± 0.54), and cold months where THI was <22 (18.39 ± 0.41). Testosterone, estradiol, superoxide dismutase (SOD), glutathione peroxidase (GPX) and lipid peroxide product (malondialdehyde, MDA) were measured in both blood and seminal plasma, while catalase (CAT) and reduced glutathione (GSH) were measured in blood and seminal plasma, respectively. Results revealed that, during the hot months, rams displayed significantly decreased testicular blood flow, increased seminal plasma MDA, decreased seminal plasma (SOD, GPx and GSH) and blood CAT antioxidant enzymes. The present study evidenced two novel findings: (a) the marked decrease in testicular blood flow volume, that is remarkable increase in both resistive index (RI) and pulsatility index (PI) values, during hot months could be negatively affected both seminal plasma enzymatic activities and seminal attributes, and (b) the SOD and GPx activities in seminal plasma of such animals were suitable predictive markers for seminal attribute evaluation.     

Quality of Fresh,Cooled, and Frozen Semen From Stallions Supplemented with Antioxidants and Fatty Acids

This study assessed the effect of oral supplementation with the primary antioxidants and fatty acids involved in spermatogenesis (L-carnitine, selenium, vitamin E, omega-3, and omega-6) on the seminal quality in fresh, cooled, and frozen semen of stallions (n = 8), using a randomized design. The animals were divided into Group I (n = 4) and Group II (n = 4) for a 30-week experiment. The two groups alternated between nutraceutical supplementation and a placebo over the course of the experiment. Semen collections were performed in two sets: once in the middle of the experiment, before the two groups switched treatments, and once at the end. The volume, appearance, sperm concentration, spermatozoa kinetics, and membrane integrity of fresh semen were evaluated. The spermatozoa kinetics and membrane integrity of cooled (for 24, 36, and 48 hours) and frozen semen were also evaluated. No differences were observed in volume, appearance, and sperm concentration between treatment and control. However, compared with placebo, nutraceutical supplementation increased (P < .05) total motility, trajectory speed, as well as plasma and acrosomal membrane integrity in spermatozoa from fresh semen. In cooled semen, nutraceutical treatment also increased (P < .05) total motility, speed, and membrane integrity of spermatozoa compared with the control. In frozen semen, supplementation increased (P < .05) spermatozoa progressive motility and plasma membrane integrity. Our results suggest a positive, synergistic effect of the antioxidant L-carnitine and selenium on spermatozoa kinetics. Similarly, the increase in plasma and acrosomal membrane integrity could be attributed to higher concentrations of polyunsaturated fatty acids (a key cell-membrane component), combined with the prevention of excess lipid peroxidation by antioxidants. In conclusion, supplementation with nutraceuticals containing fatty acids and antioxidants improved the quality of fresh, cooled, and frozen stallion semen. Therefore, nutraceutical use should increase the success of artificial insemination with cooled and cryopreserved semen.     

Changes in plasma testosterone and testicular transferrin concentration, testicular histology and semen quality after treatment of testosterone-depot plus PMSG to 3 dogs with asthenozoospermia

Three dogs diagnosed as having asthenozoospermia were given three intramuscular injections of 50 mg testosterone(T)-depot plus 250 IU pregnant mare serum gonadotropin (PMSG) at 2-week intervals, and their plasma T and testicular transferrin (Tf) concentrations, testicular histology, and semen quality were examined during the period of hormone therapy. Plasma T concentrations temporarily increased, and there was a slight improvement in spermatogenesis. Increased Tf concentrations suggested that Sertoli cell function in all three dogs was promoted by hormone treatment. The results showed that semen quality, especially the percentages of motile sperm and abnormal sperm, were improved between 1 and 5 weeks after the start of T-depot plus PMSG treatment.     

Protecting effect of vitamin E supplementation on submaximal exercise-induced oxidative stress in sedentary dogs as assessed by erythrocyte membrane fluidity and paraoxonase-1 activity

The aim of this placebo-controlled study was to investigate the effects of oral vitamin E supplementation for 10 weeks on exercise-induced oxidative damage in untrained dogs. Eight dogs were randomly assigned to a supplementation (n = 4) or control (n = 4) group and underwent two isolated submaximal exercise sessions, 10 weeks apart. Blood was collected during each session to measure erythrocyte membrane fluidity (EMF), paraoxonase-1 (PON1) activity, plasma malondialdehyde (MDA) and vitamin E concentrations. These biomarkers were measured in venous blood samples collected before (t₀), just after (t, EMF only) and 1 d (t + 1 d) and 7 d (t + 7 d) after the dogs ran on a treadmill.Prior to vitamin E supplementation, exercise induced a significant decrease in PON1 activity, EMF, vitamin E concentration and a significant increase in MDA concentration at t + 1 d. After a 10 week vitamin E supplementation period, these exercise-induced changes in PON1 activity, EMF and MDA concentration were still significant in the control group, but not in the supplemented group. These results suggested that vitamin E supplementation had a protective effect on submaximal exercise-induced oxidative damage in sedentary dogs.     

Effect of Dietary Fat Quality and Vitamin E on the Antioxidant Potential of Pigs

A randomized blocked factorial experiment was conducted with 90 young pigs. From 4 to 11 weeks of age the pigs were kept in individual pens and fed a selenium supplemented basal diet consisting mostly of propionic acid treated barley, soybean meal and dried skim milk, and containing < 0.5 mg vitamin E per kg. The treatment factors during this period were 3 dietary levels of added vitamin E (nil, 10 and 30 mg/kg) and a 6 % supplement of fresh or oxidized fat (2/3 lard and 1/3 herring oil). From 11 weeks of age until slaughter at 90 kg the pigs received the vitamin E supplements but no fat or dried skim milk. The basal diet for this later experimental period was based on untreated dry barley. Blood samples collected during the period of investigation were examined for vitamin E and for resistance against erythrocyte lipid peroxidation (ELP) in order to evaluate the antioxidant status.Analysis of variance and Student’s t-test on least squares means showed the ELP to be influenced independently by the vitamin E supplement and, during the fat feeding period, by the quality of the fat supplement, with the highest peroxidation resistance (low ELP) in the groups fed fresh fat and a high level of vitamin E. Blood vitamin E level was only influenced — positively — by the vitamin E supplement although variations in the feed vitamin E level below 10–15 mg vitamin E per kg did not result in corresponding variations in measurable blood vitamin E concentrations. In the same low range of vitamin E in the feed there was a statistically significant difference in ELP values between the different vitamin E treatment groups. No clinical manifestations of selenium-vitamin E deficiency were observed in the pigs.The ELP and the plasma vitamin E levels observed would seem to suggest that a total of 15 mg vitamin E per kg barley-based feed will not always be sufficient for growing pigs.     

High plasma estradiol-17beta levels in dogs with benign prostatic hyperplasia and azoospermia

The semen quality of 22 dogs (4 to 7 years old) with benign prostatic hyperplasia (BPH) was examined at the hospital of our university, and 4 of the 22 BPH dogs were diagnosed as azoospermic. The mean peripheral plasma estradiol-17beta (E2) level (17.3 pg/ml) of the 18 BPH dogs with spermatogenic function was higher than that of 5 normal male dogs and their mean T level (1.7 ng/ml) was lower. The mean E2 level (27.3 pg/ml) of the 4 BPH dogs with azoospermia was significantly higher than the value in the BPH dogs with spermatogenic function (P<0.01), and the mean T level (1.1 ng/ml) was significantly lower (P<0.05). Five normal male dogs were given 10 intramuscular injections of estradiol benzoate (E2B) 5 microg/kg, at 3-day intervals to investigate the relationship between high plasma E2 levels and the cause of the BPH and azoospermia. Their testes and prostates were measured and biopsied both before and 30 days after the start of E2B injections. At 30 days after the start of the E2B injections, the mean peripheral plasma T levels had decreased by half, and the mean testicular volume had decreased to 88% of original volume. The numbers of spermatocytes, spermatids, and spermatozoa in the seminiferous tubules of all of the dogs were significantly lower (P<0.05, 0.01). In addition, the mean prostatic volume increased to 130%, the mean height of the glandular epithelium decreased, and the glandular lumen became increased in diameter. These findings indicate that both BPH and serious spermatogenic dysfunction may be simultaneously induced by protracted high plasma E2 levels in dogs.     

Cholesterol Loaded Cyclodextrin Supplementation Enhances the Cholesterol-to-Phospholipid Ratio and Diminishes Oxidative Stress in Jack Spermatozoa During Cryopreservation

The present study was conducted with the hypothesis that addition of cholesterol to the extender would stabilize the sperm membranes by increasing the cholesterol-to-phospholipid (C:P) ratio and would result in an improved post-thaw semen quality and reduce oxidative stress in the jack (Martina franca) semen. Forty-eight ejaculates from six jacks were collected and analyzed for the present study. The freshly collected semen sample of each jack stallion was divided into five equal fractions after addition of the primary extender without cholesterol-loaded cyclodextrin (CLC) (C) and with 1, 1.5, 2, and 3 mg/mL CLC to obtain 120 × 10⁶ sperm/mL spermatozoa concentration. The semen was cryopreserved using customized freezing protocols. Evaluation of seminal parameters, the C:P ratio, and the oxidative status of jack spermatozoa was analyzed at all stages of cryopreservation. The oxidative status in the jack semen was evaluated by measuring malondialdehyde, glutathione and total antioxidant capacity levels. The results indicated that the mean percent values for various seminal quality parameters and the oxidative parameters were found to be significantly higher (P < .05) in CLC-treated groups with the highest values for 2 mg of CLC/120 × 10⁶ spermatozoa. In conclusion, the present study revealed that the supplementation of CLC before cryopreservation has significantly reduced the oxidative stress and also increased the C:P ratio during semen cryopreservation process. Furthermore, a reduction in lipid peroxidation levels, reduced damage to the sperm plasma and acrosome membranes and improvement in the post-thaw sperm integrity as well as stability were recorded.