The benefits of Maca (Lepidium meyenii) extract administration for male rabbits affected by environmental heat stress.

Heat stress (HS) is the most substantial environmental issue in rabbit health status impairment, which can lower productivity. Maca is a common medicinal plant with important biological activities including antioxidant, anti-inflammatory, anti-fatigue, and neuroprotective effects. The present research explored the alleged protective role of Maca extract in alleviating the adverse impact of HS on rabbits. Growing V-line 6-week-old buck rabbits (N = 48) were orally administered Maca extract at levels of 0 (MAC0), 200 (MAC2), 400 (MAC4), or 600 (MAC6) mg/head twice per week. Compared to the control groups, rabbits provided Maca had better growth performance, feed intake and feed conversion ratio. Liver and tests weights were significantly higher (p < 0.05) in Maca treated groups compared to the control. hemoglobin, white blood cell, red blood corpuscles, platelet count, hematocrit, and lymphocytes were significantly increased in the MAC4 group compared with the control group. MAC4 treatment significantly reduced aspartate aminotransferase and malondialdehyde levels compared with other groups. Moreover, the concentrations of total bilirubin, creatinine, glucose, total cholesterol, and triglycerides were lower (p < 0.05) in the MAC4 group than in the controls. Oral administration of Maca improved cortisol and testosterone values, total antioxidant capacity, and superoxide dismutase. The MAC4 group exhibited significant improvement (p < 0.05) in sperm motility, survival, membrane functionality, concentration, and libido, with a significant decrease in abnormal sperm compared with the control group. Also, aforementioned group showed sperm cells with a normal structure and intact plasma membranes, acrosomes, and well-organised axonemal components. Histopathological screening of liver sections showed moderate to severe degenerative and necrotic changes in rabbits exposed to HS, which were alleviated with Maca oral administration. In conclusion, oral administration of Maca extract at 400 or 600 mg/head-weekly enhanced growth performance, hemato-biochemical attributes, antioxidant status, and semen quality in rabbits during the hot season.

Purification of cryopreserved camel spermatozoa following protease-based semen liquefaction by lectin-functionalized DNA-defrag magnetic nanoparticles.

Although incorporating proteases into sperm medium is considered the most effective procedure to eliminate camel semen viscosity, it drastically affects viability, morpho-functional properties and, hence, fertilization potential of spermatozoa. The present work aimed at evaluating adequacy of employing magnetic nanoparticles-based sperm purification technique for eluting impaired and apoptotic camel spermatozoa from cryopreserved semen doses following protease-based semen liquefaction. Thirty cryopreserved semen doses (50 x 106 sperm/straw) representing the following liquefaction treatments: control (untreated), 0.1 mg/ml papain or 5 U/ml bromelain were used (n = 10 straws per treatment). Immediately after thawing (38°C for 40 s), sperm concentration of each straw within treatment was readjusted to 15 x 106 sperm/mL by dilution in PBS (37°C). Sperm physical and cytological properties were then assessed (non-purified semen). Thereafter, each specimen was subjected to lectin-functionalized DNA-defrag magnetic nanoparticles sperm purification, and the same sperm traits were re-evaluated after undergoing purification (purified semen). Sperm DNA fragmentation level within each group, prior to and after magnetic nano-purification, was also determined by fluorescent imaging. The results showed a dramatic improvement (p < .05) in post-thaw motility (%), viability (%), normal sperm (%), intact acrosome (%) and HOST-reacted (%) spermatozoa in protease-liquefied semen following sperm magnetic nano-purification. Additionally, the highest (p < .05) DNA fragmentation level was recorded in all cryopreserved semen groups prior to purification, whereas the lowest (p < .05) was observed in the protease-treated specimens after magnetic nano-purification. These results indicate that protease-based semen liquefaction prior to cryopreservation in conjunction with magnetic nano-purification post-thawing holds potential for reducing the proportion of damaged and dead spermatozoa, hence improving camel sperm fertilization competence.

Diluente Tris suplementado com óleo de Mauritia flexuoxa sobre a qualidade do sêmen caprino após a criopreservação / Tris diluent supplemented with Mauritia flexuoxa oil on the quality of goat semen after cryopreservation

Objetivou-se verificar os efeitos, nos parâmetros espermáticos, na integridade mitocondrial, acrossomal e de membrana em células espermáticas, desencadeados pelo uso do Tris (Tris hidroximetil aminometano) suplementado com óleo de Mauritia flexuoxacomo diluente para criopreservação de sêmen caprino. Quatro caprinos clinicamente saudáveis foram utilizados. Os animais eram alimentados diariamente com volumoso (Pennisetum purpureum, Schum.), concentrado (ração peletizada com teor de 20% proteína, 300 g/animal/dia) e sal mineral específico para Caprinos (Caprinofós®), à vontade. Dois ensaios foram realizados: I ­ Teste de toxicidade; II ­ Criopreservação do sêmen com concentrações ideais. No teste de toxicidade as concentrações avaliadas foram: 5%, 10%, 15% e 20% de diluente a base de óleo de Mauritia flexuoxa. Após o teste de toxicidade, foi escolhido a concentraçãoque apresentou o melhor resultado (5%). Logo após, foram realizadas mais 32 coletas, que foram diluídas em Tris-gema-glicerol (grupo controle) ou diluente contendo óleo vegetal (Mauritia flexuoxa). As amostras foram criopreservadas com auxílio do aparelho Tk3000®. Após o período mínimo de uma semana as palhetas foram descongeladas em banho-maria a 37 °C por 30 segundos, acondicionadas em microtubos de centrifugação e homogeneizadas para a análise imediata de motilidade, vigor espermático e morfologia. Em seguida, por meio de sondas fluorescentes foram avaliadas a integridade de acrossomo, membrana plasmática (Diacetato de Carboxifluresceína e Iodeto de Propídeo) e função mitocondrial sob microscopia de epifluorescência. Quanto a motilidade e vigor, integridade mitocondrial e acrossomal, o grupo buriti foi inferior ao grupo controle. O Tris suplementado com óleo de Mauritia flexuoxa na concentração de 5% não influenciou significativamente a qualidade espermática, porém, observou-se morfologia e integridade de membrana favoráveis. Dessa forma, sendo uma alternativa para substituição de diluentes a base de produtos de origem animal.

The objective was to verify the effects, sperm parameters, mitochondrial, acrosomal and membrane integrity in sperm cells, triggered by the use of Tris (Tris hydroxymethyl aminomethane) supplemented with Mauritia flexuoxa oil as a diluent for cryopreservation of goat semen. Four goats clinically healthy were used. The animals were fed daily with bulky (Pennisetum purpureum, Schum.), concentrate (pelleted feed with 20% protein content, 300 g / animal / day) and mineral salt Specific for Goats (Caprinofós®), ad libitum. Two tests were carried out: I - Toxicity test; II - Semen cryopreservation with ideal concentrations. In the toxicity test as selected were: 5%, 10%, 15% and 20% of Mauritia flexuoxa oil-based diluent. After the toxicity test, the concentration that showed the best result (5%) was chosen. Soon after, a further 32 samples were obtained, which were diluted in Tris-glycerol (control group) or diluent containing vegetable oil (Mauritia flexuoxa). The samples were cryopreserved using the Tk3000® machine. After a minimum of one week, the samples were thawed in a 37 ° C water bath for 30 seconds, packed in centrifugation microtubes and homogenized for immediate analysis of motility, sperm vigor and morphology. Then, by means of fluorescent probes, the integrity of the acrosome, plasma membrane (Carboxyflurescein diacetate and Propidium Iodide) and mitochondrial function under epifluorescence microscopy were evaluated. As for motility and vigor, mitochondrial and acrosomal integrity, the buriti group was inferior to the control group. Tris supplemented with Mauritia flexuoxa oil at a concentration of 5% did not significantly influence sperm quality, however, favorable motility, morphology and membrane integrity were observed. Thus, being an alternative to replace diluents based on products of animal origin.

Protocols using detomidine and oxytocin induce ex copula ejaculation in stallions.

Tricyclic antidepressives, such as imipramine, indirectly induce ejaculation by increasing the noradrenaline concentration, which triggers an α-adrenergic response, whereas α-adrenergic agonists, such as xylazine and detomidine, directly trigger ejaculation by activating the α-1 adrenergic receptors. Furthermore, serum oxytocin concentrations in stallions increase drastically before ejaculation, but decline immediately thereafter, implicating the role of this hormone in emission. The objectives of the present study were to: 1) compare the efficiency of various protocols for inducing ex copula ejaculation in stallions, 2) evaluate the benefits of including oxytocin in the protocols, and 3) compare the semen characteristics of ex copula versus in copula ejaculates. Nine protocols were used to induce ex copula ejaculation using various combinations of xylazine (X; 0.66 mg/kg, iv); oxytocin (O; 20 IU, iv), imipramine (I; 3 mg/kg, orally), and detomidine (D; 0.02 mg/kg, iv). Imipramine was given 2 h prior to the administration of α-adrenergic agonist (detomidine or xylazine) and oxytocin. If ejaculation did not occur within 10 min after treatment with an α-adrenergic agonist, a half-dose of the same product was injected. Twelve sexually mature stallions (6-26 y) were used; 9 of 12 stallions responded to the treatment. Two stallions responded to X or XO, four stallions responded to IX and IXO, one stallion responded to DO, and five responded to IDO. Stallions that responded to detomidine did not respond to xylazine. No stallion ejaculated in response to D, ID, or IO. Erections and masturbation occurred only in imipramine-treated stallions. Sperm quality was similar among all the protocols and was not significantly different from those in in copula ejaculates collected with an artificial vagina. In a separate trial, none of these protocols induced ex copula ejaculation in 2-3 y old stallions. The side effects included sialorrhea after imipramine administration in all the stallions and sedation after administration of xylazine or detomidine. In conclusion, the new protocol, IDO, and the traditional protocol, IX, had similar results, with IDO being a useful alternative protocol in stallions for which IX was not effective. Therefore, attempts using both the protocols are encouraged, as stallions that ejaculated upon administration of detomidine did not ejaculate when xylazine was administered, whereas those that responded to xylazine did not respond to detomidine.

Effects of increased levels of supplemental vitamins during the summer in a commercial artificial insemination boar stud.

Heat stress due to increasing extremes in ambient temperature and humidity results in reduced semen quality in boars. This has caused reduced efficiency of the swine industry, requiring more boars to breed the same number of sows. Vitamins such as vitamin C (VC) and E (VE) have been shown to improve semen quality in boars. Recently, vitamin D has been shown to improve semen quality in boars. The purpose of this experiment was to evaluate the effects of increased supplemental vitamins on boar reproduction during the summer season in a commercial boar stud. One hundred and sixty Pig Improvement Company (PIC) terminal line boars (n = 32 per treatment) and 39 maternal, heat-sensitive boars (n = 7 or 8 per treatment) were randomly allocated to treatment and fed a corn and soybean meal-based diet adjusted based on individual boar body condition score. A control (CNT) diet was used that met PIC recommendations for boars. Increased supplementation of specific vitamins was given in the form of a top-dress and consisted of CNT wheat middlings, CNT plus VC (560 mg/day), CNT plus 25-hydroxy vitamin D3 (VD) (125 µg/day), CNT plus VE (275 mg/day) and CNT plus VC, VD and VE (CDE). The experiment was split into three periods based on maximum daily high temperatures in the barn, where period 1 was weeks 1 to 4, period 2 was weeks 5 to 11 and period 3 was weeks 12 to 14. Semen was collected from boars as needed using the stud's normal production schedule and was analyzed for sperm quantity and quality characteristics. There were no dietary effects on semen volume, sperm concentration or total sperm production (P ≥ 0.553). Total motility of sperm was not impacted by diet (P = 0.115); although, VC tended (P = 0.064) to have a greater progressive motility than CDE. Percentages of morphologically normal sperm and normal acrosomes were not affected by dietary supplementation (P ≥ 0.157). Period effects were observed for most semen quality parameters, with quality generally becoming reduced over time. The present study demonstrates that increased supplementation of vitamins beyond PIC recommendations was not beneficial for boar reproduction during the summer.

Chrysin-induced sperm parameters and fatty acid profile changes improve reproductive performance of roosters.

Having antioxidant and androgenic effects, Chrysin was orally administrated to roosters and reproductive performance including sperm quality and fatty acid composition, testis index, fertility and hatchability rates as well as blood testosterone concentration were assessed. Twenty eight 40-week-old Ross 308 roosters were individually housed, equally divided into four groups and received different levels of capsulated Chrysin including 0 (Ch0), 25 (Ch25), 50 (Ch50) or 75 (Ch75) mg/bird/day for 12 consecutive weeks. Body weighting as well as semen and blood sampling were weekly done from 1st-10th weeks. A total of three artificial inseminations were carried out on the last two weeks of trial and collected eggs following second insemination were allotted to evaluate fertility and hatchability rates. Sperm fatty acid composition was determined using samples from 12th week. At the end of experiment (12th week) all roosters sacrificed, testis were carefully removed and testis index was calculated. Except for body weight, testis index, sperm abnormality percentage and ejaculated volume, other traits were significantly affected by Chrysin treatment. Sperm total and forward motility, plasma membrane integrity and functionality, semen concentration as well as fertility and hatchability rates were significantly higher in both Ch50 and Ch75 groups compared to control group. In spite of an increasing trend in most of n-3 and n-6 fatty acids, the n-6/n-3 ratio was significantly decreased in both Ch50 and Ch75 compared to other groups. Malondialdehyde concentration was also significantly decreased in Chrysin treated groups compared to control group. Blood testosterone level was only significantly higher in Ch75 group than that other groups. In conclusion, Chrysin administration particularly at higher levels alleviates post-peak fertility reduction in roosters; however, further research are needed to disclose involved mechanism(s).

Physiological responses and characteristics of sperm collected after electroejaculation or transrectal ultrasound-guided massage of the accessory sex glands in anesthetized mouflons (Ovis musimon) and Iberian ibexes (Capra pyrenaica).

The objective was to characterize the stress response and the seminal parameters obtained with electroejaculation (EE) or transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) in two captive but nondomestic ruminants, the mouflons and the Iberian ibex under general anesthesia. In mouflons, the physiological responses (heart and respiratory rate, rectal temperature, cortisol, creatine kinase, potassium and glucose concentrations) changed similarly with both procedures. The TUMASG procedure was faster than EE in mouflons (21.7 ± 1.4 vs. 12.4 ± 1.2 minutes, P < 0.01). In ibexes, respiratory rate, cortisol and creatine kinase concentration changes were greater with EE than with TUMASG (final respiratory rate: 62.7 ± 5.5 vs. 38.1 ± 5.6 breaths/min [P < 0.05]; final cortisol: 51.4 ± 5.1 vs. 25.3 ± 5.6 ng/mL [P < 0.001]; and final creatine kinase: 300.9 ± 99.9 vs. 87.1 ± 16.9 U/L [P < 0.001]). Electroejaculation provided better results in some sperm parameters (mouflons: sperm score: 3.4 ± 0.3 vs. 2.6 ± 0.2 [P < 0.01]; total number of sperm ejaculated: 982.4 ± 299 vs. 710.0 ± 542.2 [P < 0.05]; ibexes: sperm with progressive motility: 47.7 ± 6.2 vs. 20.5 ± 8.3 [P < 0.05]). The transrectal ultrasound-guided massage of the accessory sex glands appears to be an alternative technique to collect sperm from wild ruminants, reducing the need for electrical stimuli and thus decreasing the undesired responses of EE in the more sensitive species. On the other hand, better fresh sperm may be collected with EE. However, TUMASG provides practical advantages in animal welfare, firstly in these wild species more sensible to stress management and capture myopathy.

Effects of dietary supplementation with astaxanthin and ß-carotene on the semen quality of goldfish (Carassius auratus).

This study was conducted to investigate the effects of two carotenoids (astaxanthin and ß-carotene) on the sperm quality of goldfish Carassius auratus (Linnaeus, 1758). For this purpose, six diets containing concentrations of 50, 100, and 150 mg/kg of synthetic astaxanthin and ß-carotene were added to a basic carp diet. One group of fish was also fed with a control diet (no added carotenoids). Osmolality, spermatocrit value, and sperm concentration significantly increased in the treatment supplemented with 150 mg/kg of astaxanthin (296.6 ± 1.1 mOsm/kg; 29.2 ± 0.6%; 17.2 ± 0.4 × 10(9) cells/mL, respectively) and ß-carotene (295.2 ± 2.1 mOsm/kg; 32.5 ± 1.6%; 17.9 ± 0.5 × 10(9) cells/mL, respectively). The highest concentration of astaxanthin (10.4 ± 1.4 mg/kg) was recorded in the treatment of A150 (P < 0.05) and did not differ between ß-carotene treatments. The highest motility was observed in the A150 and B150 treatments, and the lowest was observed in the control group (P < 0.05). The artificial fertilization of the treated males with the similar females (fed with the control diet) showed that the fertilization rate in the A150 treatments was higher than in the other treatments (P < 0.05). In conclusion, dietary supplementation with 150 mg/kg of astaxanthin improves osmolality, motility, fertilization rate, and sperm concentration.

Alcohol diluent provides the optimal formulation for calcium chloride non-surgical sterilization in dogs.

BACKGROUND: Surgical castration is widely used to sterilize male dogs, but has significant impacts on time to perform the operation, recovery of the animals as well as cost, which can limit population control programs. Previous research has shown intratesticular injection of calcium chloride dihydrate (CaCl2) in saline to be a promising alternative to surgery. However, long-term azoospermia was not maintained at dosages low enough to avoid side effects. In the search for an optimized formulation, the current investigation is the first study on long-term sterilization effects of intratesticular injection of CaCl2 in either lidocaine solution or alcohol in dogs. CaCl2 at 20% concentration in lidocaine solution or alcohol was administered via intratesticular injection to groups of 21 dogs each. The treated animals were examined at 2, 6, and 12 months for sperm production, blood levels of testosterone, and side effects; at time zero and 12 months for testicular size and semen volume. The experimentally treated animals were compared to a control group receiving saline injection only. RESULTS: Testicles of dogs treated with CaCl2 in either diluent significantly decreased in size. After administration of CaCl2 in lidocaine solution, sterility was achieved for at least 12 months in 75% of treated dogs. However, optimal long-term contraceptive effectiveness was achieved with CaCl2 in alcohol, which resulted in azoospermia over the 12-month study period. Testosterone levels significantly decreased following treatment with CaCl2, and sexual activity disappeared. Although testosterone returned to baseline levels by 12 months for the group treated with CaCl2 in lidocaine, dogs injected with CaCl2 in alcohol had a 63.6% drop in testosterone level, which remained at the low end of physiological range throughout the study. No adverse effects were noted. CONCLUSIONS: A single, bilateral intratesticular injection of 20% CaCl2 in 95% ethanol was a reliable method for induction of sterilization in 18-28 kg male dogs in this study. The approach showed long-term efficacy and reduced sexual behavior. This chemical method of sterilization might provide an effective, efficient alternative to surgical castration that can have positive impacts on dog welfare.

A dose-finding, long-term study on the use of calcium chloride in saline solution as a method of nonsurgical sterilization in dogs: evaluation of the most effective concentration with the lowest risk.

BACKGROUND: Canine overpopulation is a global issue with serious health and welfare implications. Nonsurgical methods of sterilization could yield positive impacts on this problem, but no long-term data on such methods are available. The objective of the current investigation was to determine the effects of intratesticular injections of calcium chloride dihydrate (CaCl2) in saline in dogs over a one year period. Five concentrations (0%, 10%, 20%, 30%, 60%) of CaCl2 in saline were administered via intratesticular injection to groups of 10 dogs each. Total sperm count and motility, blood levels of testosterone, and side effects were examined at 0, 2, 6, and 12 months post-injection (PI). Testicular size and semen volume were examined at 0 and 12 months PI. RESULTS: Total sperm count, semen volume and testosterone showed significant dose-dependent decreases upon treatment with 10%-60% CaCl2 compared with either the control group (0% CaCl2) or baseline for each treatment group. Azoospermia was achieved for at least 12 months PI in 60% and 80% of treated dogs after administration of a 10% and 20% CaCl2, respectively. Treatment with 30% or 60% CaCl2 resulted in azoospermia in 100% of dogs, but more side effects were observed, while no side effects were noticed at lower doses. For each treatment group, testosterone levels had decreased an average of 35%-70% at 6 months following treatment. However, testosterone levels rebounded by the 12-month time point in all groups except the highest dosage group (60% CaCl2), which remained at the low end of physiological range throughout the study. Sperm motility dropped to zero or near zero in all dogs treated with CaCl2. Testicular size was significantly smaller at 12 months PI for all groups when compared to baseline. CONCLUSIONS: This first long-term study confirms reports of the efficacy of CaCl2 sterilization. However, at dosages free of adverse events, calcium chloride in saline may not provide permanent sterilization as previously believed. Future work should explore optimized solvents to increase the permanence of the well-tolerated 20% formulation.

Study of sperm concentration, seminal plasma composition and their physiological correlation in the Persian sturgeon, Acipenser persicus.

The objectives of the present study were to determine ionic and organic composition of seminal plasma, sperm concentration and their relationships in the Persian sturgeon (Acipenser persicus). In this regard, ionic content (Na(+) , K(+) , Cl(-) , Ca(2+) and Mg(2+) ) and organic content (total protein, glucose, cholesterol and triglyceride) along with sperm concentration were measured in 17 specimens of the Persian sturgeon. The seminal plasma contained 59.53 ± 2.56 mm/l sodium, 9.1 ± 1.42 mm chloride, 4.72 ± 0.3 mm potassium, 1.45 ± 0.075 mm calcium and 0.7 ± 0.072 mm magnesium. The following organic contents were found: total protein 0.11 ± 0.02 g/dl, glucose 22.18 ± 4.16 mg/dl, cholesterol 6.67 ± 1.04 mg/dl and triglyceride 15.2 ± 0.65 mg/dl. The mean sperm concentration was estimated to be 1.6 ± 0.12 (×10(9)  sperm/ml). A significant relationship was found between sperm concentration and K(+) of seminal plasma (r = 0.533, p < 0.05). Significant correlations were observed between ionic contents: Na(+) vs Cl(-) (r = -0.854, p < 0.01) and Mg(2+) vs K(+) (-0.583, p < 0.05). Also, level of triglyceride was negatively correlated with Mg(2+) (r = -0.503, p < 0.05). Presented data could be considered as a complementary study for developing special extenders and protectant solutions for improving artificial fertilization in this valuable species.

The detoxication of nitrate by two antioxidants or a probiotic, and the effects on blood and seminal plasma profiles and reproductive function of New Zealand White rabbit bucks.

Forty-two New Zealand White male rabbits were housed individually in wire cages and randomly distributed among six experimental groups of seven rabbits each, during 16 to 61 weeks of age. There were three main nitrate groups: 0 (tap water), 350 and 700 ppm. Within the 700 ppm of nitrate, there were four subgroups, in which one group was used as control group and the other three groups were supplemented with either 200 ppm of ascorbic acid (vitamin (Vit) C), 200 ppm of Vit E with 0.2 ppm of selenium (Se) and 1000 ppm of probiotic. The nitrate was supplemented as a sodium nitrate. The aim is to test the ability of Vit C and Vit E, Se and probiotic on the deleterious effects (blood and seminal plasma biochemical constituents, semen quality and productive performance) of nitrate in drinking water. Rabbits given nitrate at 700 ppm had significantly lower plasma globulin, red blood cells (RBCs), hemoglobin (Hgb), packed cell volume % (PCV%) and total antioxidant capacity (TAC) than those given the other concentrations of nitrate. Vit C, Vit E with Se and probiotic resulted in significantly (P < 0.05) greater Hgb, RBCs, PCV% and TAC than those of bucks given water supplemented with only 700 ppm nitrate, but the aspartate aminotransferase and alanine aminotransferase concentrations in seminal plasma were lower. Testosterone in the blood plasma and the seminal plasma was significantly (P < 0.05) lower in rabbits given 700 ppm nitrate than in those given other concentrations of nitrate. Vit C, Vit E with Se and the probiotic significantly increased testosterone, fertility, number of offspring and total offspring weight of rabbits sired by bucks supplemented with 700 ppm of nitrate.

The effect of individualized homeopathic treatment on the semen quality of bulls with reproductive disorders: a case series.

BACKGROUND: Poor semen quality of pedigree bulls has major economic implications in cattle breeding. AIMS: To evaluate the effect of homeopathy on the semen of bulls with reproductive disorders. METHODS: The behavioral, clinical and spermatic characteristics of four Nelore bulls were evaluated. The bulls received individualized homeopathic treatment mixed into the feed and administered once per day. Semen was collected using an artificial vagina. Successful collection, freezing, effective doses, and the number of doses of semen per collection were compared before and after homeopathic treatment. RESULTS: The bulls were treated with Sulphur, Phosphorus, Lycopodium and Argentum nitricum all in 30CH dilution. The rates of successful collection were 97.14%, 100%, 96.67% and 30.77% pretreatment and 95.45%, 100%, 94.67% and 96.77% at post-treatment for bulls A, B, C and D, respectively. The average number of doses per ejaculate pre and post-treatment were 102.67 (SD 74.41) vs. 300.08 (SD 180.58), 0 vs. 234.78 (SD 96.12), 0 vs. 105.12 (SD 54.98), 0 ± vs. 107.37 (SD 52.12) respectively. Many of these differences were statistically significant. CONCLUSION: The use of homeopathy apparently improved the production of viable doses of semen from bulls with previous freezing problems and poor semen quality. Controlled studies should be conducted.

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.

Suplementação de touros com sabões cálcicos de ácidos graxos poli-insaturados e qualidade seminal pré e pós-congelação / Bulls supplemented with calcium soaps of polyunsaturated fatty acids and semen quality before and after freezing

O objetivo deste experimento foi avaliar a qualidade seminal de touros suplementados com sabões cálcicos de ácidos graxos poli-insaturados (PUFA). Vinte touros das raças Angus, Brangus, Hereford e Braford foram distribuídos aleatoriamente em dois grupos, cada um com dez animais. Cada grupo recebeu uma dieta composta de volumoso, ração comercial, sal mineral e um de dois suplementos: funcional com a incorporação de sabões cálcicos de PUFA ou energético, com a incorporação de raspa de mandioca (Manihot esculenta, Crantz). Durante o período experimental, a qualidade seminal foi avaliada utilizando testes rotineiros (volume, concentração, motilidade em massa, motilidade individual e avaliação da morfologia espermática) e testes complementares (teste hiposmótico, coloração dupla giemsa/tripan-azul e teste de termorresistência). O sêmen in natura do grupo de touros suplementados com sabões cálcicos de PUFA comercial (Megalac-E®) apresentou aumento na motilidade espermática (83,3% vs. 75,3%), na percentagem de espermatozoides com acrossoma íntegro (94,8% vs. 91,8%) e na percentagem de espermatozoides viáveis (98,0% vs. 96,6%) quando comparado com o sêmen do grupo de touros suplementados com raspa de mandioca (P<0,05). O tipo de suplemento não afetou o volume seminal, a concentração espermática e concentração de testosterona em sangue (P>0,05). Foram encontradas interações entre tratamento e número de coleta para percentagem de espermatozoides morfologicamente normais e percentagem de espermatozoides positivos ao teste hipo-osmótico (P<0,05). Após a descongelação, o tipo de suplemento não afetou a motilidade dos espermatozoides, o teste hipo-osmótico e o teste de termoresistência rápido (P>0,05). A suplementação com sabões cálcicos de PUFA aumentou as percentagens de espermatozoides com acrossoma íntegro (48,0% vs. 39,2%) e de espermatozoides com membrana plasmática integra (51,5% v.s 42,2%) pós-descongelação.

The objective of this experiment was to evaluate semen quality of bulls supplemented with calcium soaps of fatty acids polyunsaturated. Twenty Angus, Brangus, Hereford and Braford bulls were randomly allocated into two groups, each with ten animals. Each group received a diet consisting of forage, commercial ration, mineral supplements and supplement. Two supplements types were evaluated: functional supplement, with fatty acids polyunsaturated (PUFA) incorporation or energy supplement, with cassava meal (Manihot esculenta, Crantz) incorporation. During trial period the semen quality was evaluated using routine test (volume, concentration, mass motility, individual motility, sperm morphology) and complementary test: (hypo-osmotic test, giemsa/tripan-azul coloration and thermotolerance test). The semen of group bulls supplemented with commercial calcium soaps of PUFA (Megalac-E®) showed greater sperm motility (83.3% vs. 75.3%), percentage of sperm with intact acrosome (94.8% vs. 91.8%) and percentage of sperm with intact plasma membrane (98.0% vs. 96.6%) when compared with semen of group bulls supplemented with cassava (P<0.05). The supplement type did not affect semen volume, sperm concentration and testosterone concentration in blood (P>0.05). There were interactions between treatment and collection period for percentage of normal sperm and percentage of spermatozoa with hypo-osmotic positive test (P<0.05). The type of supplement did not affect sperm motility, the hypo-osmotic test and fast thermotolerance test after thawing (P>0.05). PUFA calcium soaps supplementation increased percentage of spermatozoa with plasma membrane integrity (51.5 vs. 42.2%) and spermatozoa intact acrosome (48.0% vs. 39.2%) after thawing.

Effect of maca supplementation on bovine sperm quantity and quality followed over two spermatogenic cycles.

Maca (Lepidium meyenii Walpers), is an Andean crop that grows between 3,800 and 4,500 m a.s.l. The persistent interest in this plant is based on its assumed effects on fertility of male mammals due to the prevalence of certain, partially specific, secondary compounds. The present study aimed at evaluating the effect of maca supplementation on quality and quantity of semen, mating behavior, and clinical status of peripubertal breeding bulls. The experiment followed a cross-over design lasting for 23 wk with 3 wk of adaptation and baseline measurements, and 2 x 10 wk of treatment feeding thus covering two times the complete 8-wk spermatogenic cycle. Seventy-eight 55 wk to 84 wk old breeding bulls received either no maca (control) or maca (233 mg dried hypocotyls/kg body weight/day) for 10 wk followed by 10 wk without maca (maca early) or maca only in the last 10 wk (maca late). Measurements were always made in the last 2 wk of each period. Apart from standard analyses, ejaculates were analyzed by flow cytometry. Data was evaluated by analysis of variance considering the repeated measurement structure of the data. Significant treatment by measurement period indicated direct or carry-over effects of maca. Maca supplementation had no direct effect on body weight, testes circumference, rectal temperature, mating behavior, and ejaculate volume. However, supplementing maca in the first 10 wk period increased the number of sperms in the second 10 wk period, i.e., when the animals no longer received maca. The DNA fragmentation index and the visually assessed motility of the sperms of bulls, that initially showed a borderline sperm quality, were significantly improved with early maca supplementation, while no such effect was observed in the two other groups. No effects occurred in the proportion of intact sperm plasma membranes or acrosomes or both. In conclusion, maca supplementation seems to improve sperm quantity and quality of bulls to a certain degree, while mating behavior appears unaffected.

Organic selenium enhances the antioxidative status and quality of cockerel semen under high ambient temperature.

1. The objective was to examine the effect of supplemental dietary organic selenium (Se) on improving semen quality and antioxidative status in male domestic fowls exposed to high ambient temperature. 2. Thirty-six Egyptian local cross males, 42 weeks old, were housed individually in cages in an open-sided building under 16 h L:8 h D and were provided with commercial feed and water ad libitum. In the house average daily temperature ranged from 33 to 36 degrees C and relative humidity from 60 to 70%. Birds were divided into 4 experimental treatments (n = 9) and were fed the basal diet supplemented with 0 (control), 01, 02 or 03 mg organic Se/kg in the form of a yeast source (Sel-Plex) for 8 weeks. 3. Under heat stress conditions, inclusion of organic Se in the cockerel diets enhanced the semen quality traits, including the sperm count and motility and reduced the percentage of dead sperms in a dose-dependent manner. 4. In seminal plasma, organic Se supplementation ameliorated some of the adverse effects of heat stress on lipid peroxidation and antioxidative properties. The inclusion of organic Se (03 mg/kg) in the cockerel diets doubled seminal plasma glutathione peroxidase (GSH-Px) activity, compared to controls and reduced lipid peroxidation. 5. In conclusion, supplemental dietary organic Se improved semen quality characteristics when cockerels were subjected to heat stress. It increased both sperm count and motility, reduced the percentage of dead sperm and enhanced the antioxidative status of seminal plasma.

Effects of dietary vitamin supplementation and semen collection frequency on reproductive performance and semen quality in boars.

The present study was undertaken to assess the relevance of increasing the daily provision of dietary vitamins on vitamin metabolic status and semen characteristics of boars under controlled and commercial conditions as well as to evaluate the efficiency of this vitamin supplement to allow boars to cope with intensive semen collection frequency. In the first experiment, 39 boars were allocated to 2 dietary treatments, a basal diet (control) and the basal diet supplemented with extra fat- and water-soluble vitamins (Vit). Within each treatment, boars were submitted to 2 regimens of semen collection frequency: 3 times per 2 wk (3/2) and 3 times per week (3/1) over a 12-wk period. Afterwards, all boars were intensively collected (daily) for 2 wk. A resting period of 4 wk followed, and all boars were collected 2 times per week. Thereafter, collection frequencies were reversed, and the same procedure was followed until the end of the intensive collection period. A second experiment was conducted in commercial conditions at a commercial stud, and 252 boars were randomly allocated to the control and Vit dietary treatments. All boars were collected 2 times per week over a 6-mo period. Classical measurements of ejaculate and sperm quality were assessed, and blood samples were collected throughout both experiments to quantify vitamin concentrations. In the first experiment, vitamin concentrations in blood and seminal plasma increased in Vit boars (P < 0.05); however, vitamin concentrations were not affected by collection frequency (P > 0.14). The Vit supplement did not affect sperm production or sperm quality (P > 0.28), although semen volume increased during the 12-wk periods for Vit boars (P < 0.05). The 3/1 boars produced fewer doses per ejaculate than 3/2 boars (P < 0.01); however, the cumulative sperm production for the 12-wk periods increased by 19% in 3/1 boars compared with 3/2 boars. In the second experiment, blood plasma concentrations of vitamin B(9) were greater (P < 0.01) in Vit than control boars. The vitamin supplement did not increase sperm production of boars (P > 0.61). In conclusion, dietary supplements of fat- and water-soluble vitamins increase the amount of vitamins available for the animal, and the collection frequencies had no effect on vitamin status. Moreover, in spite of an effect on the ejaculate volume, the dietary supplement of extra vitamins had no effect on sperm production or quality.

Quality and lipid composition of spermatozoa in rabbits fed DHA and vitamin E rich diets.

The effects of fish oil (FO) and vitamin E (vE) dietary supplementation on semen quality, sperm susceptibility to lipid peroxidation, tocopherols content and fatty acid profiles were studied in rabbits. Fifty-two rabbit bucks randomly divided in four groups received a control diet and enriched diets containing either FO (1.5%, w/w), vE (200mg/kg) or both. Semen volume, concentration, motility and viability were analysed at various time-points and the lipid composition was assessed on sperm cells. The phospholipid fatty acid profile was determined: n-6 PUFA were the major fatty acids found, with a proportion of 42%, whereas the n-3 PUFA accounted for nearly 1%, mainly represented by C22:6n-3 (docosahexaenoic acid, DHA). FO supplementation produced a seven-fold increase in the content of DHA in sperm phospholipids and a comprehensive rearrangement of the phospholipid fatty acid composition, while an unexpected negative effect of feeding high level of vE on the proportion of total PUFA was found. Despite the remarkable changes observed in sperm lipid composition, semen quality parameters were not affected by the dietary treatments and the interaction between the two dietary supplements had a significant effect only on sperm concentration. An increase in semen production by ageing and a concomitant rise in sperm susceptibility to in vitro peroxidation was found. alpha- and delta-tocopherol, present in rabbit sperm in similar amount, were not affected by dietary treatment. delta-tocopherol content had a significant linear negative regression with age and showed a significant negative correlation with the susceptibility to peroxidation values.

Effect of semen dilution to low-sperm number per dose on motility and functionality of cryopreserved bovine spermatozoa using low-density lipoproteins (LDL) extender: comparison to Triladyl and Bioxcell.

Artificial insemination with doses containing low-sperm numbers has been utilized to optimize the use of elite bulls. Hen egg yolk is widely used as a cryoprotective agent in semen freezing extender protecting the spermatozoa. Its action is due to the presence of low-density lipoproteins (LDL) in the hen egg yolk. The objectives of the present study were to evaluate the effects of the semen dilution to low-sperm number/dose on sperm motility and integrity of sperm plasma membrane in the cryopreservation process, using two commercial extenders (Triladyl, Bioxcell and LDL extender prepared in our laboratory, 97% purity. Fifteen ejaculates were collected from five fertile crossbred bulls (Bos taurusxBos indicus). After collection, sperm motility was examined by Computer-Assisted Semen Analysis (Hamilton Thorne), morphological sperm characteristics were evaluated by differential interference microscopy and the integrity of plasma membranes was determined using the hypo-osmotic swelling test. The semen was subsequently divided into three aliquots and diluted with the three extenders into 120 x 10(6), 60 x 10(6) and 20 x 10(6)sperm/mL, corresponding to 30 x 10(6), 15 x 10(6) and 5 x10(6) sperm/dose, respectively. This study revealed that LDL extender was more effective in preservation of motility and integrity of the plasma membrane of spermatozoa than Bioxcell and Triladyl (p<0.05), but no significant difference was observed between Triladyland Bioxcell. Therefore we can conclude that LDL extender could be used instead of Triladyl or Bioxcellat low semen concentration per dose for elite bulls, it also could be envisaged for the industry of sex-stored semen.