Therapeutic effects of vitamin E supplementation in 4 dogs with poor semen quality and low superoxide dismutase activity in seminal plasma

Four dogs with poor semen quality, low seminal plasma superoxide dismutase (SOD) activity and low blood plasma testosterone (T) levels were orally administered one vitamin E tablet containing 50 mg α-tocopheryl acetate per dog daily for 4 weeks. The mean values of semen quality were temporarily improved after the start of vitamin E treatment and the values of 4, and 5 weeks after that were significantly different from those before the treatment (P<0.05–0.001). The mean blood plasma T and seminal plasma SOD activity values slightly increased in the 4 dogs after the treatment. The results of the present study indicate that poor semen quality in dogs with low seminal plasma SOD can be improved by vitamin E treatment.     

高温对荷斯坦种公牛精液品质及精清生化指标的影响研究

以荷斯坦种公牛为试验牛研究了高温对其精液品质和精清生化指标的影响。结果表明:(1)高温可造成种公牛精液品质显著下降。使原精活力、精子密度、活精子百分数和顶体完整率分别比春季下降3·13%(P<0·05)、34·8%(P<0·01)、15·0%(P<0·01)和17·8%(P<0·01),精子畸形率比春季上升25·5%(P<0·01)。(2)夏季荷斯坦种公牛精清睾酮含量仅为4·31pg/mL,极显著低于非高温季节(P<0·01)。(3)高温环境使得荷斯坦种公牛精清钾、钙、镁处于最低水平(P<0·01),精清碱性磷酸酶(ALP)、酸性磷酸酶(ACP)处于最高水平(P<0·01),使精清钠由春季至夏季呈显著下降趋势。     

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.     

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.     

Influence of long-term treatment with equine somatotropin (EquiGen) on gonadal function in stallions with poor semen quality

The aim of the present study was to investigate the spermatogenic and Leydig cell activity in stallions with impaired semen quality after treatment with equine somatotropin. Experiments were performed using 18 adult clinically healthy stallions with poor semen quality which did not pass breeding soundness evaluation. The animals were randomly divided into a treatment (n = 9) and a control (n = 9) group. Over a period of 90 days, nine stallions received a daily intramuscular injection of 10 mg recombinant equine somatotropin (EquiGen, BresaGen Limited, Adelaide, Australia) and 9 control animals were injected with the same amount of physiological saline solution. During and until 2 months after treatment, semen characteristics and daily sperm output as well as plasma testosterone concentrations were determined monthly in all stallions. In addition, testosterone concentration measurement after stimulation with hCG was performed in all animals immediately before and at the end of the treatment period as well as 2 months later. Our results demonstrate that equine somatotropin (EquiGen) given daily in a dose of 10 mg per animal during 90 days had no significant effect neither on plasma testosterone concentrations and hCG-induced testosterone release nor on semen quality parameters in adult stallions with poor semen characteristics.     

Effect of organic and inorganic selenium supplementation on semen quality and blood enzymes in buffalo bulls

The present study aimed to evaluate the effect of organic and inorganic selenium (Se) supplementation on semen quality and blood serum profiles of buffalo bulls. Nine mature buffalo bulls were divided into three groups: control (non‐supplemented); organic Se (10 mg Sel‐Plex®/head twice weekly) and inorganic Se (10 mg sodium selenite/head twice weekly). Semen was collected twice a week for 3 months during Se supplementation. Semen properties were evaluated from fresh ejaculate. Moreover, fructose concentration, aspartate and alanine transaminase (AST and ALT) activities, total protein and total cholesterol were assayed in seminal plasma. Additionally AST, ALT, testosterone and Se levels were determined in the blood serum. Results showed that Se supplementation significantly (P < 0.05) influences the semen parameters during 3 months of treatment. Organic Se significantly (P < 0.05) increased the percentage of viable sperms compared to inorganic Se and the control group. Fructose concentration was significantly higher (P < 0.05) in the seminal plasma of organic Se‐treated bulls. Serum testosterone and Se concentrations were significantly (P < 0.05) increased in the Se supplemented groups than the control group. In conclusion, Se supplementation improved the parameters of buffalo bull semen and more precisely, organic Se was more effective for the improvement of semen quality and some blood components than inorganic Se.     

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.     

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.     

Effects of finasteride on size of the prostate gland and semen quality in dogs with benign prostatic hypertrophy

OBJECTIVE: To determine the effect of the 5alpha-reductase inhibitor finasteride on prostatic diameter and volume, semen quality, and serum dihydrotestosterone (DHT) and testosterone concentrations in dogs with spontaneous benign prostatic hypertrophy (BPH). DESIGN: Double-blind placebo-controlled trial. ANIMALS: 9 dogs with BPH. PROCEDURE: Five dogs were treated with finasteride for 16 weeks (0.1 to 0.5 mg/kg [0.05 to 0.23 mg/lb] of body weight, PO, q 24 h); the other 4 received a placebo. Prostatic diameter, measured radiographically, prostatic volume, measured ultrasonographically, semen quality, and serum DHT and testosterone concentrations were evaluated before and during treatment. After receiving the placebo for 16 weeks, the 4 control dogs were treated with finasteride for 16 weeks, and evaluations were repeated. RESULTS: Finasteride significantly decreased prostatic diameter (mean percentage decrease, 20%), prostatic volume (mean percentage decrease, 43%), and serum DHT concentration (mean percentage decrease, 58%). Finasteride decreased semen volume but did not adversely effect semen quality or serum testosterone concentration. No adverse effects were reported by owners of dogs in the study. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that finasteride can be used to reduce prostatic size in dogs with BPH without adversely affecting semen quality or serum testosterone concentration.     

Biochemical observations on beagle dog semen.

Semen samples were collected at weekly intervals for six weeks from eight sexually mature beagles previously shown to produce normal ejaculates. Seminal plasma and sperm fractions were separated by centrifugation and the sodium, potassium, alanine and aspartate aminotransferases, acid and alkaline phosphatase concentrations in the two fractions determined. Regression analysis of the mean weekly values obtained from physical and biochemical examination of the ejaculates showed that sodium ion concentration was highest in seminal plasma. The highest levels of aminotransferases were found in sperm fractions. Those enzymes may be indices of abnormal or damaged spermatozoa. Acid and alkaline phosphatase activity was 100 times greater in seminal plasma than in sperm fractions. Phosphatase concentrations are likely to be dependent on prostate activity. Measurement of acid phosphatase in canine semen therefore may be a useful index of prostate function. The motility of the semen samples was independent of the potassium concentration in seminal plasma. However, there was some evidence of a correlation between sperm motility and the enzyme and sodium content of seminal plasma.     

Performing a complete canine semen evaluation in a small animal hospital

Normal dog semen ranges in volume from 1 to 30 mL per ejaculate and contains 300 million to 2 billion sperm, of which more than 70% are progressively motile and morphologically normal. Dog semen should contain fewer than 10,000 bacteria per mL; higher numbers are indication of an infection of the male reproductive tract and usually are associated with cytologic evidence of inflammation (neutrophils in semen sediment) and with decreased progressive motility and decreased numbers of morphologically normal sperm. Measurement of pH of seminal plasma of dogs with infection of the reproductive tract may provide information on determining the choice of antibiotic. Measurement of seminal plasma alkaline phosphatase in azoospermic dogs is an indicator of tubular patency to the level of the epididymides.     

The influence of centrifugation on quality and freezability of stallion semen

The aim of the present study was to investigate the influence of various centrifugation methods on sperm loss and quality of frozen-thawed semen. From at a total of 8 Warmblood stallions of the National Stud Farm in Avenches, 3 ejaculates each were collected and seminal plasma was removed using 3 different centrifugation regimes. In method I (reference method) centrifugation occurred by a speed of 600 x g during 10 minutes. In method II 1000 x g was used during 2 minutes while in method III centrifugation was performed by 2000 x g during 2 minutes. After centrifugation 90%, of the supernatant was removed and sperm loss calculated. After resuspension of the pellet with freezing medium, functional membrane integrity was evaluated by HOS-test and motility determined. In frozen-thawed semen motility, viability as well as functional membrane integrity (HOS-test) and acrosome status using chlortetracyclinassay (CTA) were assessed. Our results demonstrate that mean sperm loss (I, 1.9%; II, 8.7%; III, 3.7%) was significantly (P < 0.05) different between the three centrifugation regimes. Regarding semen quality of frozen-thawed semen, HOS in method III (52.1%) was significantly lower than in methods I (55.5%) and II (55.3%). Evaluation of the acrosome status by CTA showed that more than 70% of sperm cells were capacitated and 25% capacitated and acrosome reacted. From our results we conclude that sperm loss and functional membrane integrity (HOS-test) in frozen-thawed semen were significantly influenced by the centrifugation regime. Therefore, stallion semen should be centrifuged at 600 x g during 10 minutes before freezing in order to obtain low sperm loss and a good quality of frozen-thawed semen.     

Seasonal variation in semen quality of bulls and correlations with metabolic and endocrine parameters

Sperm output and semen quality of 17 bulls sampled over 12 months showed minimal output in mid-winter and late summer and minimum quality in late summer. Monthly measurements of luteinising hormone and testosterone concentration in plasma and testosterone concentration in semen were made over 12 months. Serum non-esterified fatty acids (NEFA), albumin and total protein were also measured for the final seven months of this period. Plasma testosterone showed a strong negative correlation with sperm numbers two months hence but not in the current month. Plasma testosterone by bull and semen testosterone by month was also correlated with sperm output. Plasma luteinising hormone three months and serum total protein two months prior was positively correlated with sperm numbers ejaculated and the normality of sperm morphology, possibly by affecting the luteinising hormone dependent A0 to A1 spermatogonial division. NEFA was correlated with initial and post freezing/thawing motility in the current month, possibly by affecting membrane stability. The value of examining the bull in diagnosing infertility of cows where nutritional stress may have occurred is suggested, as is the use of albumin/total protein and NEFA measurements as a prognostic aid for time to return to normality of function of such bulls.     

Effects of herbal preparation on libido and semen quality in boars

The objective of this study was to investigate the effects of a preparation from herbal extracts (PHE) on libido and semen quality in breeding artificial insemination boars. Ten fertile boars were divided into control and experimental groups according to significant difference of libido. There were no differences in semen quality between groups. Animals were fed a commercial feeding mixture for boars. The feeding mixture for the experimental group was enriched with PHE, which was prepared from Eurycoma longifolia, Tribulus terrestris and Leuzea carthamoides. Duration of the experiment was 10 weeks. Samples of ejaculate were collected weekly. Libido was evaluated according to a scale of 0-5 points. Semen volume, sperm motility, percentage of viable spermatozoa, sperm concentration, morphologically abnormal spermatozoa, daily sperm production and sperm survival were assessed. Amounts of mineral components and free amino acids were analysed in seminal plasma. Significant differences were found in these parameters: libido (4.05 ± 0.22 vs 3.48 ± 0.78; p < 0.001), semen volume (331.75 ± 61.91 vs 263.13 ± 87.17 g; p < 0.001), sperm concentration (386.25 ± 107.95 vs 487.25 ± 165.50 × 10(3) /mm(3); p < 0.01), morphologically abnormal spermatozoa (15.94 ± 11.08 vs 20.88 ± 9.19%; p < 0.001) and Mg concentration (28.36 ± 11.59 vs 20.27 ± 13.93 mm; p < 0.05). The experimental group's libido was increased by 20% in comparison with the beginning of the experiment. Results of this study showed positive effect of PHE on libido and some parameters of boar semen quality.     

Seasonal variations in semen quality of bulls: correlations with environmental temperature

Seventeen bulls were used for a 12-month survey of semen quality and for the estimation of plasma progesterone and testosterone and semen testosterone concentrations. Sperm output showed two minima, in mid-winter and late summer, and the percentage of abnormal sperm was highest and their ability to survive freezing was lowest at the summer minimum. Plasma progesterone concentration was negatively correlated with the total ejaculate content of testosterone, and positively with the local average maximum daily temperature. Temperature showed a quadratic relationship with the percentage of abnormal sperm ejaculated one month later, with the minimum percentage occurring at 14.5 degrees C. Temperature also showed a quadratic relationship with the numbers of sperm in semen ejaculated two months later, with the maximum number occurring at 17 degrees C. These relationships may reflect the impaired or enhanced survival of the temperature sensitive meiotic prophase and alterations in the output of testosterone and progesterone by the testis.     

Effect of dietary vitamin E and selenium supplementation on semen quality in Cairn Terriers with normospermia

Among others, selenium (Se) and vitamin E (VitE) have been provided to dogs to improve semen quality. However, scientific evidence documenting an effect in dogs is lacking. The aim of this study was to investigate the effect of supplementation of these antioxidants on various ejaculate parameters in a randomized, double‐blinded trial using Cairn Terrier males exhibiting normal seminal quality parameters. Three dogs each were fed a standardized diet and supplemented with 0.1 mg Se, 100 mg VitE or 0.1 mg Se + 100 mg VitE/dog for 3 months. Ejaculate analyses (volume, progressive motility, vitality, morphology, concentration) were performed before inclusion (D0) and after 1, 2 and 3 months (+1, +2, +3). At the same time, glutathione peroxidase (GSH‐PX) and VitE in seminal plasma (SP) and GSH‐PX in blood samples were determined. Vitamin E levels in SP were below the detection limit (1.0 mg/L) in all samples. GSH‐PX in blood (164.0–2794.4 IU/L) and SP (18.4–4326.0 IU/L) was highly variable. Supplementation only significantly affected the total percentage of sperm head abnormalities (p = .011). Time significantly affected the percentage of morphologically abnormal sperm (p = .025), sperm head abnormalities (p = .007), proximal droplets (p = .001) and GSH‐PX in SP (p = .015). Additionally, a significant interaction between time and group was identified for the percentage of membrane‐intact sperm (p = .048), head abnormalities (p = .018), acrosomal defects (p = .043) and proximal droplets (p = .002). Although some effects could be identified for selected parameters, we failed to identify a clear trend about how a 3 months VitE and/or Se supplementation affects semen parameters in normospermic Cairn Terriers.     

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。     

Changes in plasma testosterone levels and semen quality after 3 injections of a GnRH analogue in 3 dogs with spermatogenic dysfunction

Three male Beagles with low plasma luteinizing hormone and testosterone levels and spermatogenic dysfunction (SD-dogs) were given 3 weekly subcutaneous injections of a gonadotropin-releasing hormone analogue (Buserelin; GnRH-AB). The plasma T level of all of the SD-dogs increased and peaked at 4-6 weeks after the first injection. The total number of sperm and the sperm motility of the three SD-dogs increased 5-7 weeks after the first injection. Therefore, the authors concluded that 3 weekly injections of GnRH-AB transiently improves semen quality in some cases of canine spermatogenic dysfunction.     

Effect of vitamin supplements on some aspects of performance, vitamin status, and semen quality in boars

The aim of the present study was to determine the effects of dietary supplements of vitamins on vitamin status, libido, and semen characteristics in young boars under normal and intensive semen collection. Sixty Landrace, Yorkshire, and Duroc boars were allocated randomly from 6 to 10 mo of age to one of the following diets: 1) basal diet (industry level) for minerals and vitamins (Control, n = 15); 2) basal diet supplemented with vitamin C (ASC, n = 15); 3) basal diet supplemented with fat-soluble vitamins (FSV, n = 15); and 4) basal diet supplemented with water-soluble vitamins (WSV, n = 15). After puberty (approximately 12 mo of age), semen was collected at a regular frequency (three times every 2 wk) for 5 wk. Thereafter, all boars were intensively collected (daily during 2 wk). A recovery period (semen collection three times every 2 wk) followed and lasted for 10 wk. Sperm quality (percentage of motile cells and percentage of morphologically normal cells) and quantity (sperm concentration, semen volume, and total sperm number) were recorded as well as direct and hormone related measurements of boar libido. Blood and seminal plasma samples were taken to monitor vitamin status. High concentrations of B6 (P < 0.05) and folic acid (P < 0.05) were observed in the blood plasma of WSV boars, whereas greater concentrations of vitamin E (P < 0.01) were obtained in FSV boars. In the seminal plasma, folic acid concentrations tended to be greater in WSV boars (P < 0.08). During the intensive collection period, there was a tendency (P < 0.06) for semen production to be greater in WSV boars, the effect being less pronounced (P < 0.10) in FSV boars. During the recovery period, the percentage of motile sperm cells was greater in WSV boars (P < 0.03) and, to a lesser extent, in FSV boars (P < 0.10) compared with Control boars. Sperm morphology and libido were not affected by treatments. These results indicate that the transfer of vitamins from blood to seminal plasma is limited and the dietary supplements of water-soluble and fat-soluble vitamins may increase semen production during intensive semen collection.     

Does Coenzyme Q10 Exert Antioxidant Effect on Frozen Equine Sperm?

During semen cryopreservation, the sensitivity of equine sperm to oxidative stress is increased by the eliminated seminal plasma. Thus, antioxidant addition to the semen extender can be helpful to the sperm survival after freezing and thawing. This work aimed to test whether coenzyme Q10 (CoQ10) added in different concentrations to the INRA 82 freezing extender has antioxidant function on equine sperm to improve its fertilizing ability. Semen samples from five stallions were frozen with the extenders: (T1) INRA 82, control, (T2) T1+ 5 μM CoQ10, (T3) T1+ 25 μM CoQ10, and (T4) T1+ 50 μM CoQ10. After sample thawing, sperm motility and kinetics characteristics were evaluated using a computer-assisted sperm analysis and sperm membrane functionality and integrity were evaluated with a hypo-osmotic swelling test and an epifluorescence microscopy, respectively. The nitrite (NO₂^-) and hydrogen peroxide (H₂O₂) concentrations of the semen samples were measured with spectrophotometry. There was no difference on the sperm characteristics among all treatments (P > .05). However, the 25 μM CoQ10 (T3) decreased NO₂⁻ concentration (6.7 ± 2.2 μM/μg protein) compared with the treatments T1, T2, and T4 (64.3 ± 3.7, 59.4 ± 5.3, 45.1 ± 8.6 μM/μg protein), respectively, as well H₂O₂ concentration (1.8 ± 0.3 μM/μg protein) compared with the control (4.6 ± 0.4 μM/μg protein) and 5 μM CoQ10 treatments (4.8 ± 0.2 μM/μg protein, P < .05). In conclusion, 25 μM CoQ10 plays a significant role as antioxidant to the frozen equine sperm, decreasing NO₂⁻ and H₂O₂ concentrations. Thus, its addition to the INRA 82 freezing extender may be beneficial to the fertilizing ability of equine semen.