جستجوی مقالات مرتبط با کلیدواژه « sperm » در نشریات گروه « علوم دام »

The present study was conducted in order to use different levels of curcumin in the diluting sperm epididymis of Shal breed rams during storage at five degrees Celsius in the form of a completely randomized design with four treatments in six replications. Sperm samples were extracted from the testicular tissue and used in the Tris-egg yolk diluent. Different concentrations of curcumin including 0, 10, 25 and 50 μM were added to the diluents. The samples were gradually cooled at 5°C and after stabilization at this temperature, in 6, 12, 24 and 48 hours, the parameters of total motility, progressive motility, viability, membrane integrity and the percentage of sperm abnormalities were evaluated. The results showed that the use of 25μM curcumin improved the parameters of total motility, progressive motility, viability and membrane integrity only at 6 and 12 hours, but no significant difference was observed at 24 and 48 hours compared to other treatment groups. Regarding sperm abnormalities, no significant difference was observed between the treatments in all the examined times. The results of this study indicate that using a concentration of 25μM curcumin can lead to the improvement of ram epididymal sperm quality in the initial periods of storage, but it loses its effect in longer periods of time. Therefore, it is recommended to use nanotechnology methods to improve the effectiveness of curcumin, especially during long-term storage of ram sperm.

Sperm cryopreservation has become an indispensable tool in reproductive biology. However, maintaining fertility in frozen sperm is very important. The aim of this study is to investigate the effects of selenium nano-particles on the storage of dog semen at 4 ˚C during a period of 48 hours. In this study, 20 ejaculates were collected from 4 mix dogs and diluted in a Tris-based diluent. Then, they were divided into 4 parts in control and treated with 0.5, 1 and 1.5 µg selenium nano-particles groups. Sperm parameters including evaluation of total and progressive motility sperm, motility characteristics and viability of sperm were evaluated for 48 hours. The obtained results showed that in concentrations of 1 µg [progressive sperm (27.03±1.48), total motility (49.48±1.27) and viability (41.40±1.03)] and 0.5 µg [progressive sperm (30.76±1.67), total motility (53.18±1.54) and viability (43.15±1.58)] of selenium nanoparticles compared to the control group, but in the concentration of 1.5 µg was lower than the control group (p< 0.05). As a result, the present study showed that adding selenium nanoparticles in concentrations of 0.1 and 0.5 µg to dog semen can improve the parameters of dog semen after liquid storage, but in concentrations of 1.5 µg it causes toxic effects and overall agitation. And it reduces sperm viability.

 Rooster’s reproductive performance is an indispensable component of breeder production because it plays a vital role in the maximum production of fertilized eggs. Existence feed supplement in the poultry industry, intermediary metabolites have been including in the diet to improve fertility and reproductive outcomes. Carnitine (β-hydroxy-γ-trimethylaminobutyrate), vitamin-like-amino acid, is a quaternary ammonium compound, that has multifunctional roles in reproduction. High concentrations of L-carnitine (LC) are present in epididymal lumen, where it participates in sperm energy balance and the maturation of spermatozoa. In light of previously reported breeder birds supplemented with dietary LC have shown improvements in semen traits and fertility parameters. The present study is an attempt to investigate the effects of several levels of dietary LC supplementation on semen quality parameters and gonadosomatic and hepatosomatic indexes at maturity and production peak.

For the present experiment, thirty-six Ross (12-week-old) breeder broilers were used for 22 weeks in a completely randomized design with three treatments (0, 250 and 500 mg L-carnitine in kg of diet) and twelve replications. All roosters were fed standard isocaloric (2754 kcal/kg) and isonitrogenous diet (12 % protein). The birds for 22 weeks in a completely randomized design with three treatments (0, 250 and 500 mg / kg of LC in the diet) and six replications were used. During the adaptation period (21-24 weeks of age), the roosters were trained by abdominal massage for semen collection. After the experimental period was commenced (24 weeks of age), semen samples were collected and evaluated for seminal attributes every two weeks (from week 24 to week 34). The following parameters were determined immediately after the semen collection; to measure the semen samples were collected weekly to evaluate semen volume, total motility, membrane functionality, mitochondria activity parameters. Also, to determine gonadosomatic and hepatosomatic indexes at 24 and 34 weeks of age, six birds in each treatment weighing then were slaughtered and immediately the testes and liver were removed and weighed.

The highest sperm motility (96.60%) was observed in birds fed 250 mg LC (P <0.04). By increasing the level of LC in the diet, sperm membrane functionality improved linearly (6.8% increase compared to control) (P <0.04). A linear trend (P = 0.06) was observed in mitochondrial activity with increasing levels of LC in the diet (69.31, 72.00 and 76.25). LC plays an essential role in energy metabolism by carrying over fatty acids in the mitochondria matrix for β-oxidation producing energy. Therefore, it provides a better supply of energy for spermatogenesis and normal physiology of sperm, are presumably improved by an optimum level of LC, as a result, sperm concentration and live. These data provide evidence that LC can be effectively used in diets up to 500 mg/kg of diet or 30 mg/kg of body weight /day for semen improvements of rooster’s breeder. At 24 weeks of age, the changes in gonadosomatic index (0.55, 0.68 and 0.64) were affected by different levels of LC (P <0.01). During testis development in the chicken (from 2 to 15 weeks of age), there is no significant increase in testicular weight, however, the early stage is the most important period for testicular development. The mature testis has seminiferous tubules with a multilayered epithelium representing the different stages of spermatogenesis. Sexual maturity is associated with the highest testes weight and consequently with the highest plasma concentration of reproductive hormones. Gonads of the mature male broiler breeder are organized into separate, comfortably discernible cellular correlations and functional compartments. It has been accepted that steroid hormones biosynthesis and generation of spermatozoa are two major actions that the testicles fundamentally carry out. The improvements in gonadosomatic Index of roosters observed in this study in response to dietary LC may be attributed, at least partly, due to improved utilization of dietary nitrogen, achieved through more efficient fat oxidation by LC. Testicles contain the seminiferous tubules and the interstitial space. Seminiferous tubules are the functional elements of the testis and sertoli cells are the principal structural basis of the seminiferous epithelium, inhabiting on the substratum membrane.

The addition of 250 and 500 mg of L-carnitine to the diet due to the increase in gonad index led to an improvement in sperm quality parameters at the beginning of the production period (puberty).

The need for artificial insemination in the ewe is to access fresh and frozen sperm. Cryopreservation is a suitable method for long-term storage of semen and sperm, but may cause a minor irreversible damage to the sperm cell, especially its membrane region (Khorramabadi et al., 2017). The oxidation of fatty acids leads to the production of free oxygen radicals (ROS). These radicals are necessary in normal conditions for certain physiological activities and sperm processes, but excessive production of ROS in the sperm can reduce membrane fluidity, DNA fracture, damage to proteins, and ultimately reduced sperm motility and fertility. (Bakhshayesh et al., 2017). Silymarin, the scientific name of Silbum marianum, is the English name Milk Thistle and is a potent inhibitor of oxidative stress as a potent antioxidant. Due to the nature of its friend fat, silymarin is firmly attached to the plasmid membrane compounds, thus preventing fracture and its collapse by increasing membrane strength. Increasing the amount of active oxygen species and lipid peroxidation disrupts mitochondrial membrane, decreases ATP and damage to sperm acesomes, which ultimately reduces the progression of sperm motility. (Kvasnikova et al., 2003). The mechanism of action of salimarin is through stimulation of the ribosomal RNA and protects the membrane from oxidative damage. They also stated that silymarin stimulates the activity of the antioxidant enzymes of superoxide dismutase (SOD) and glutathione peroxidase (Wellington and Jarvis, 2001) . Silymarin improves the sperm motility and survival, sperm abnormalities, and preservation of the sperm membrane after freezing-thawing. Improvement of semen quality in both freezing and cooling is due to the strong silicomain antioxidant capacity (Fakorzai et al., 2008; Longpyram et al., 2013). .

Regarding the anti-oxidant effects of silymarin, this study aimed to investigate the effect of silymarin on storage of ram sperm.

In this research, four rams of pure ghee were used 2-3 years old and each ram was 6 times sperm. Sperm was performed twice a week by artificial vagina. After the sample was transferred to the laboratory, a series of preliminary evaluations including sample size, sample color, wave motion, total mobility, in-situ motion, progressive motion and live weight, and abnormal sperm and density were performed. If standards were met, Necessary (scaling over 2.5 billion sperm and progressive movement above 70%) dilution was performed with treatments. The diluents were prepared prior to sampling and placed in a bin Marie temperature of 37 ° C. Tris-based diluent was used to contain 2.73 g of tries, 1.4 g of fructose, 1 g of citric acid and 100 mg streptomycin in 100 ml sterile distilled water. To prepare the diluent, 73% of the prepared solution will be mixed with 20% egg yolk and 7% glycerol. In order to dilute the sprays from trace based diluent, egg yolk, semen collected with diluent without antioxidants (control) and silymarin 5 and 10 μg / ml mix After cooling for 90 minutes in the refrigerator and reaching 5 ° C, it was placed in 4 cm above the liquid nitrogen for 8-10 minutes and then immersed in liquid nitrogen. The post-mortem examinations of sperm quality properties included liveliness, total motion, progressive motion, integrity of the plasma membrane (tests) on days 0, 15, 30 of the experiment.

According to the results of the experiment, the diluents containing the levels of silymarin significantly increased the total motion, progressive, live survival of ram sperm after the freeze-thawing process compared to the control group Were opened. Addition of different concentrations of silymarin was 5 and 10 μg with mean of 74.45% and 67.33%, respectively, compared to the control group, significantly increased the total sperm motility with mean of 68.07 and 60.95%, significantly Progressive motility of frozen ram sperm were frozen (p <0.01). According to the results, the in situ movement in the control group (5.92 ± 0.92) was lower than that of the 5 and 10 μg silymarin (6.38 ± 0.92) and (8.9 ± 0.92) respectively In the least amount.

Application of silymarin as an antioxidant in the ram sperm diluent system reduces oxidative damage and improves sperm quality properties such as mobility, viability and the health of acrosome and plasma membrane during freezing process. Was opened. Adding 5 μg of silymarin to diluted ram minus 10 μg and control group reduced fat peroxidation and also improved the structure and performance of sperm during storage.

The low fertility rate of frozen sperm of poultry compared to other species is a serious challenge for artificial insemination in commercial flocks. This challenge may be related to some physiological characteristics of poultry sperm. On the other hand, osmotic, chemical and mechanical stresses during cooling processes are the most important reasons that cause structural, biochemical and functional changes in sperm, which results in a series of cascade phenomena that will lead to a decrease in sperm fertility. Quercetin is a bioactive compound with a plant origin, which has a polyphenolic chemical structure and has antioxidant properties. This composition, as an important flavonoid and a strong antioxidant, eliminates free radicals and reduces oxidative stress in chicken spermatogonia. The purpose of this study is to use quercetin in nano form in the diluent of rooster sperm during storage at 4 degrees.

Immediately after collecting semen from roosters, initial evaluations were done. Semen collected from all 15 roosters were mixed together and added to the dilution medium in order to eliminate individual effects. Different concentrations of quercetin (10, 15 and 20 μM) in three forms (unprotected and inside liposomal and NLC structures) added to base diluent composition. Then the diluted semen samples were gradually cooled at 4°C for 3 hours. Then, at time zero (immediately after the samples reach the temperature of 4°C), 24 and 48, the motility parameters, membrane integrity, lipid peroxidation, the percentage of abnormal sperms and the percentage of viability of the samples were evaluated.

The results indicated that the experimental treatments did not have a significant effect on the evaluated parameters at zero time. During 24 hours of storage at 4 degrees, 15 μM of quercetin loaded in NLC significantly improved the parameters of sperm total motility, progressive motility, viability and integrity of sperm membrane compared to the control group. During 48 hours of storage at 4 degrees, the treatment of 15 μM quercetin loaded in NLC caused a significant improvement in the parameters of sperm total motility, progressive motility, viability and integrity of sperm membrane compared to the control group. Also, the results showed that the experimental treatments did not have a significant effect on the abnormality percentage of rooster sperm during storage at 4 degrees in three times of 0, 24 and 48 hours. During 24 and 48 hours of storage at 4 degrees, the treatment of 15 μM quercetin loaded in NLC caused a significant decrease in sperm MDA compared to the control group.

The results of this experiment show that the treatment of 15 μM quercetin loaded in NLC improves many evaluated parameters and therefore it is recommended for use in rooster sperm diluent.

Unlike evaluation of male fertility which is done based on the phenotype records of females e.g., conception rate, semen traits are directly measured in bulls. The objective of this study was to identify genomic regions associated with sperm traits in Holstein bulls using the genome wide association based on gene set enrichment analysis.

For this purpose, phenotype records included 1508 genotyped Holstein bulls were used for ejaculate volume, sperm concentration, number of motile sperm, progressive sperm motility, and number of progressive motile sperm in each ejaculation. The evaluation of genome-wide association was carried out PLINK software (v. 1.90). In the next step, using the biomaRt2 R package R the SNP was assigned to genes if they were within the genomic sequence of the gene or within a flanking region of 15 kb up- and downstream of the gene. For the assignment of the genes to functional categories, the GO, KEGG, DAVID and PANTHER databases were used.

In this research, 11 SNP markers on chromosomes 1, 2, 5, 7, 8, 15, 18, 20, 21, 25, and 27 associated with SPEF2, SEPT12, SLC24A, BSP3, ETNK1, ERBB4 (sperm motility and number of progressive motile sperm), DSCAML, MYH3, RPM1 , RPM2, CAPSL (number of sperm and sperm concentration), DMRT1, and PPARγ genes (ejaculate volume) were identified. Some of these genes in the significant regions were consistent with some previous studies. In pathway analysis, 15 pathways from gene ontology were associated with the sperm traits. Among those pathways, the purine nucleotide metabolic process, cellular calcium ion homeostasis and regulation of fertilization biological pathway had an important role in the progressive sperm motility and number of progressive motile sperm. Also, the calcium channel regulator activity, PPAR signaling pathway, steroid hormone mediated signaling pathway and focal adhesion had significant association with ejaculate volume and sperm concentration.

The Identification of these candidate genes and selection bulls using genomic selection can considerable to improve male fertility and benefits of animal breeding centers and decrease production prices for consumers.

During sperm freezing, the production of reactive oxygen species and the reduction of antioxidant activity cause damage to sperm. Most of the stress is due to the high production of reactive oxygen species and the impermeability of antioxidants to mitochondria. In this study, the effects of four levels of a combination of 2, 4- dinitrophenol which is a targeted mitochondrial antioxidant and Luteolin antioxidants, which are among the strongest flavonoid compounds with antioxidant and antimicrobial properties, were evaluated in lecithin-based Lake Extender.

For this purpose, semen samples from 15 roosters were collected by dorso-abdominal massage method. Semen samples are mixed together after initial evaluation after diluting and adding different levels, treatment 1 (0.5 nanomolar 2, 4- dinitrophenol + 0.75 micromolar Luteolin), treatment 2 (0.5 nanomolar 2, 4- dinitrophenol + 1 micromolar Luteolin), treatment 3 (0.75 nanomolar 2, 4- dinitrophenol + 0.75 micromolar Luteolin) and treatment 4 (0.75 nanomolar 2, 4- dinitrophenol + 1 micromolar Luteolin), of antioxidants, samples were frozen by nitrogen vapor. After thawing, the samples were evaluated for sperm performance parameters.

The results of the present study showed that the addition of treatments 2 and 3 to the leak diluent during freezing and thawing of rooster sperm significantly increased the parameters of total motility and progressive movement and also treatment 3 significantly increased sperm velocity in a straight line and the criterion of direct sperm motility. Also, treatments 2 and 3 significantly increased survival compared to the control group and reduced the amount of semen malondialdehyde and sperm with abnormal morphology during freezing and thawing (p<0.05). Also, treatment 3 was able to increase membrane integrity. Plasma increase compared to the control group (p<0.05).

The results of the present study show that the combined use of these two antioxidants has increased the properties of both antioxidants and improved the quality and quantity of sperm.

The study aimed to investigate the effect of asparagus extract on quality semen of Arabian rams at different storage times at a temperature of 4degrees Celsius. Sperm collection was performed from 10 Arab rams with an average age of 2.5 years per week (once a week) for six weeks. Afteradding 0, 1.5, 3 and 4.5% asparagus extract to the semen diluent, sperm quality parameters were evaluated at 0, 24, 48, and 72 hours after storage inliquid condition. At zero hour, sperm viability in diluent had 3% asparagus extract was higher than diluent containing 0 and 4.5% extract (P<0.05).After 24 hours of semen storage, the parameters of total motility, progressive motility, plasma membrane integrity and viability were higher in spermdiluent containing 1.5 and 3% of the extract (P<0.05). Qualitative parameters of sperm in 48 hours after sperm collection and storage in diluentcontain 3% of extract had significant and better performance (P<0.05). The highest rate of plasma membrane integrity, progressive motility and totalsperm motility was observed in diluent containing 3% extract in 72 hours after sperm storage (P<0.05). According to the results of this study, the useof asparagus extract in the diluent at the rate of 3% improves the sperm quality of Arabian rams after cooling conditions.

Long-term storage of semen is essential for achieving the benefits of artificial insemination (Tuncer et al., 2010). This is carried out by sperm cryopreservation, which stops the sperm metabolic activities, allowing long storage (Bailey et al., 2000). This process affects the sperm quality (Wang et al., 1991) by introducing mechanical and chemical damage, production of reactive oxygen species, oxidative stress, and reducing antioxidant activity (cysteamine is an aminothiol antioxidant as an effective scavenger). Cysteamine is known to have been reported in some studies to improve the freezing of ram sperm (Bucak et al., 2007). The aim of the present study was to determine the antioxidant effects of cysteamine on the functional parameters of cystemine in Lake Extender based on soybean lecithin.

This study was carried out in University of Tabriz research station. For this purpose, 15 adult roosters of 25 weeks were used. Sperm collection was done by dorsal-abdominal massage. The roosters were habituation for one month and sperm collection was performed twice a week. First, sperm were examined for volume, concentration and color, and only samples with volume of 0.2 to 0.7 ml and motility greater than 80% were used. To eliminate the individual effects, the confirmed samples were pooled. Four levels containing control, 15, 30, and 45 μM cysteamine were then added to Lake Extender containing 1/4 glycerol and sperm added to the extender containing different levels of antioxidant. The cooling process was carried out in two steps. The samples were adjusted to 4°C and after two hours the samples were cooled to 4°C. Then, samples were transferred to the refrigerator for one hour more, then they were drawn into 0.25 ml straws, placed 4 cm above nitrogen vapor for 7 min, and then immersed in liquid nitrogen. They were stored in liquid nitrogen until carrying out the tests. For thawing, the straw containing the samples was removed from the tank and immersed in water for 30 seconds at 37°C in water bath. The motility parameters were evaluated using CASA, viability by Eosin-Nigrosin staining, membrane integrity by Host tests, sperm abnormality by Hancock test and lipid peroxidation by MDA.

Addition of cysteamine at all levels significantly improved motility parameters. At the 30 and 45 μM levels total motility significantly increased and 30 μM level improved progressive motility, VAP, VSL and VCL parameters (P <0.05). The results show that the addition of cysteamine amino acid improves sperm quality. Addition of 30 μM level significantly increased viability and plasma membrane integrity of rooster sperm (P <0.05); Also significantly decreased malondialdehyde compared to the control group (P <0.05). The addition of cysteamine reduced sperm abnormality which was not significant. Discussion Freezing causes oxidative stress and induces eversible damage such as reduced viability and fertility (Najafi et al., 2014). This study was conducted to evaluate the addition of cysteamine as a supplement to counter oxidative damage during freezing and thawing (Blesbois, 2011). Normally, the antioxidant system in the semen eliminates the free radicals in the semen which are disturbed by cryopreservation process (Safa et al., 2017). Adding antioxidants can control ROS production (Amini et al., 2015). Antioxidants in different species and at different doses have shown different results. Addition of cysteamine to rooster spermatozoa during freezing and thawing significantly increased motility and survival parameters, which was in agreement with the results of Bucak et al., (2007) reported that the addition of cysteamine to ram sperm improved sperm motility parameters (Najafi et al., (2014) also reported an increase in motility and viability of ram sperm with addition of 2, 4 and 6 mM cysteamine levels, but at 8 mM level these parameters were significantly reduced. This was probably due to the high dose of diluent used. But it was in contrast to the study of Thananurak et al., (2019) who reported that adding levels of 0.001, 0.002, 0.004 and 0.01 decreased motility and viability. Low levels of cysteamine induce cysteine to enter the cell, producing glutathione as an intracellular antioxidant. Cysteamine at high doses produces large amounts of hydrogen peroxide, causing oxidative stress and reducing glutathione peroxidase activity (Besouw et al., 2013). According to reports (Partyka et al., 2013) cysteine increases motility and viability of rooster sperm. Glutathione peroxidase is one of the enzymes with peroxidative activity that plays an important role in sweeping high oxidations and protecting the cell from oxidative stress. Glutathione can also restore the oxidized vitamins E and C and restore them to the original antioxidant structure (Almasi et al., 2014). Cysteamine as a glutathione synthase can play a major role in reducing free radicals. In the present study, the amino acid cysteamine decreased malondialdehyde concentration and increased plasma membrane integrity and normal sperm during the freezing-thawing process, which is in agreement with the report of Najafi et al., (2014) which reported that addition of 6 mM reduced the concentration of malondialdehyde in ram semen compared with the control group. Not only does cysteamine improve sperm quality after freeze-thawing, but also it increases sperm resistance during artificial insemination in the reproductive female tract (Najafi et al., 2014). In the present study, cysteamine did not significantly decrease the percentage of abnormal sperm compared to control group.

Antioxidants have different results depending on the doses which are used and the treated species, and by understanding the antioxidant mechanism and achieving the optimal dose, the best results can be obtained. In the present study, the use of cysteamine amino acid as an antioxidant supplement improved sperm functional parameters and significantly reduced lipid peroxidation at level 45 μM.

Using 36 Hubbard grandparent roosters at 45 weeks of age, the present study investigated the effects of supplementing vitamin E and soybean lecithin in the roosters' diet on their semen quality. The experiment was conducted in a completely randomized design with four treatments including basal diet, basal diet supplemented with vitamin E (300 mg/kg), basal diet supplemented with lecithin (1%), and basal diet supplemented with lecithin & vitamin E. Roosters were fed their diets for 60 days and semen collection was performed on days 0, 20, 40 and 60 of the experiment. The results showed that the treatment containing vitamin E + lecithin significantly improved total sperm motility and sperm concentration (P<0.05). The semen volume and sperm kinematic values were not affected by experimental diets (P>0.05). Although experimental treatments had no effect on abnormal morphology of sperm and DNA fragmentation, but membrane integrity and sperm viability were significantly increased in roosters fed diet supplemented with soybean lecithin and vitamin E (P<0.05). Malondialdehyde (MDA) concentration was also shown to be significantly lower in treatment containing vitamin E and soybean lecithin (P<0.05). The interaction effects of time× diet on total and progressive motility, sperm concentration, viability, membrane integrity and MDA content were significant (P<0.05) and these traits improved in the last days of the experiment due to diet supplemented with vitamin E+ soybean lecithin. Overall, it was concluded that supplementing aged roosters' diet with vitamin E and lecithin appears to improve sperm quality traits.

 Reproductive performance of domestic animals has a great impact on the profitability of the farms. It has been shown that several factors such as breed, age, season and nutritional management affect the quality of the produced semen and consequently fertility of rams. Among various nutritional factors influencing semen quality, fat has a great impact on both quantity and quality of produced spermatozoa so that its value is correlated with cell membrane fluidity, potent intracellular signal transduction molecules, and susceptibility to oxidative damage. It has been postulated that by participating in sperm plasma membrane fatty acids (FA), the ingested fat can change the ratio of polyunsaturated to saturated fatty acids (PUFA: SFA) and thereby improve several aspects of sperm quality. Also, lipids comprise a wide-range class of molecules that not only is served as a source of energy but also play a crucial role in the structure and function of spermatozoa. Dietary n‑3 PUFA supplementation has also shown to improve semen quality parameters in rams. Flaxseed oil contains up to 90% PUFAs, of which about 50% is α-linolenic acid. Several studies conducted during past decades indicate that dietary flaxseed supplementation improves sperm parameters of different species such as bovine, goat and rabbit.

Materials and Methods :

Fifteen mature 3-5 years old Kurdish ram weighing 65±2.5 kg (mean ±SE) were randomly allocated into three groups during 12 weeks of the experimental period. Animals were individually fed a standard basal diet supplemented with different levels of flaxseed. Treatment included FLS-0 (basal diet; control), FLS-5 (basal diet containing 5% flaxseed) and FLS-10 (basal diet containing 10% flaxseed. Blood and fresh semen samples were collected at weeks 1 and 12 of the experiment. The collected samples were examined for sperm concentrations, sperm motility, viability, acrosome integrity, host test and fatty acids profiles. The testis circumference was measured with flexible cloth tape. The largest circumference of the testes and both scrotum was measured after pushing the testes firmly into the scrotum. To measure plasma concentrations of glutathione peroxidase and testosterone, blood samples were collected from the jugular vein of all the rams at the beginning and after 1 and 12 weeks of feeding experimental diet.

Results and Discussion:

 Flaxseed supplementation did not affect testicles circumference, however, supplementation of flaxseed increased plasma concentrations of testosterone in (10% flaxseed) FLS-10 group compared to (5% flaxseed) FLS-0 (P<0.05). Treatment, time and their interaction did not affect glutathione peroxidase (GPX) activity. Semen concentration, proportion of live sperm, total motility and plasma membrane functionality was higher in FLS-10 compared to FLS-5 and FLS-0 groups (P<0.05). Flaxseed supplementation tended to increase percentage of sperm with normal acrosome (P<0. 01); however, percentage of abnormal was decreased in FLS-10 compared to the other corresponding groups (P<0.05). Treatment, time and interaction effect of treatment × time did not affect LIN, STR, ALH and VCL (P>0.05). However, BCF and VSL were improved in FLS-10 group as compared to the FLS-5 and FLS-0 group (P<0.05). A significant interactive effect of treatment × time was noted for VAP, where flaxseed supplementation did not affect VAP at first week of the experiment but higher VAP was recorded in FLS-10 compared to FLS-0 and FLS-0 groups after 12 weeks of flaxseed feeding (P<0.05). Flaxseed supplementation did not affect percentage of myristic, palmitic, stearic, oleic, linoleic, EPA and DPA, but the proportion of DHA, and the ratio of n-3:n-6 PUFA and SFA:PUFA was affected by the treatments (P<0.05). Dietary inclusion of flaxseed increased proportion of DHA and n-3: n-6 PUFA ratio and decreased SFA: PUFA ratio in FLS-10 compared to FLS-5 and FLS-0 (P<0.05). The interactive effect of treatment × time on DHA, n-3: n-6 and SFA: PUFA ratio revealed that there was no significant effect between treatment after first week of the experiment; however, proportion of DHA and n-3: n-6 PUFA ratio was higher and SFA: PUFA ratio was lower in FLS-10 compared to FLS-5 and FLS-0 (P<0.05). the significant correlations between addition of 10% flaxseed and most of the evaluated semen characteristics including live sperm, total motility, plasma membrane functionality, and acrosome status following 12 weeks of treatment feeding was in agreement with previous findings in male goat and rabbit. Other fatty acids concentrations, such as, linoleic acid, and docosahexaenoic acid (DHA) was improved by dietary flaxseed supplementation.

Conclusion:

 It can be concluded that adding 10% flaxseed to the Kurdish ram diet out of the breeding season can improve sperm quality. Sperm fatty acid composition can also be affected by dietary fat. But more research is needed to look at the effects of other flaxseed products, such as oil and powder. It is recommended that similar studies be conducted during the breeding season and with other amounts of flaxseed.

Mitochondria as sperm energy supplier motors have unique properties including antioxidant non-permeability, so the ROS concentration in this organelle is 5 to 10 times more than that of the plasma membrane. Therefore, the researchers have designed an antioxidant which can cross the mitochondrial membrane and is able to inhibit ROS in this organelle to minimize the oxidative damage. Targeted antioxidant 4, 2-dinitrophenol has the ability to cross the mitochondrial membrane. The purpose of this study was to add different levels of targeted antioxidant to semen to purify mitochondria from reactive oxygen species to reduce oxidative damage and improve sperm motility and viability. In this study, 8 Ghezel rams (2 years old) were used for semen collection by artificial vagina in early spring. After initial evaluation, the samples with normal properties were pooled and after dilution process, 0, 0.25, 0.5, 0.75, and 1 nM 4, 2-dinitrophenol were added to the samples and frozen after 2 hours of cooling. After one month, samples were thawed and evaluated for motility parameters, viability, membrane health, sperm abnormality and lipid peroxidation. Results showed that addition of 0.5 nM of the antioxidant significantly increased motility parameters and decreased abnormal sperm (P <0.05). Also suplimentation of 0.5, 0.75 nM 4, 2-dinitrophenol significantly increased viability and membrane integrity and reduced the amount of lipid peroxidation (P <0.05).

Freezing is one branch of cryobiology knowledge that process to protecting and long term keeping of cell in very low temperature. Sperm freezing commonly done in infertility treatment centers and keeping sperm banks and livestock industry. Sperm can survive decrease out of body and environment temperature. freezing is so important one of influential destructive factors in reducing duration of sperm storage is creation of ROS. It needs very low, but it is increasing has destructive effects on sperm (lenzi et al. 1994). Use of antioxidants in sperm extender have better results to deal with these reactive oxygen species. It is believed that ROS induces lipid peroxidation in the sperm membrane and The peroxidation of the resulting fatty acids has toxic effects on the sperm that Leading to a decrease in sperm function (Sanuka and Corps 2004). Antioxidants are compounds that control, stop and neutralize intermediates derived from the energy production process in the electron transport chain of cells (aerobic activity) known as active oxygen species, and in a group of free radicals (Zinee et al. 2000). Adding antioxidants to semen diluent during freezing of sperm improves the quality and improvement of semen parameters such as motility, viability and acrosome membrane integrity (Mironoksuk et al. 2018). Selenium is an essential micronutrient element to human health Which is toxic at high concentrations. Selenium is a constituent of selenoproteins that they have an enzymatic and structural role in biochemistry. Selenium is the main component of selenoenzymes in the center of all of these proteins, there is the amino acid of selenocysteine Which acts as an oxidation-reducing agent. However, one of the limiting factors for selenium consumption is bioavailability (the rate of entry into circulation and tissues) and its toxicity (Tarz et al. 2007). Therefore, the use of other forms of selenium that is less toxic and more beneficial is more appropriate. Selenium nanoparticles are low toxicity compounds and bioavailable and effectively increase the production of selenoproteins in the body (Peng et al. 2007). Selenium is a cofactor or activator of the glutathione peroxidase enzyme Which is one of the strongest antioxidants in the body and catalyzes the decomposition of lipid peroxides and hydrogen peroxide (Hill et al. 1996). The glutathione peroxidase enzyme destroys free radicals within the cytoplasm (Ozball et al. 2008). Typically, the concentration of most of the elements in the liver is higher than the other organs, but in the case of selenium, it has the highest concentration in the testis. High concentration of selenium in testis indicates the protective role of this element and its related enzymes in spermatogenesis (Schumberger et al. 1983).

Semen was collected with an artificial vagina twice a week from 4 selected adult Ghezel rams during non-reproductive season. Ejaculates were immediately evaluated primarily for parameters including volume, concentration, motility, viability and abnormal sperm. And if they had the required standards (high concentration of 3 billion sperm per ml, motility above 70% and volume greater than 0.5 ml), they were selected as suitable samples for dilution. The semen was diluted with extender with no antioxidants (control) and containing 1 µg/ml Nano Selenium and 2 µg/ml Nano Selenium and then immediately aspirated to 0.25 ml freezing straw. After recording, treatments were packaged and refrigerated in 5°C for 1.5 hours. Then, the packaged straw containing semen were placed on 4 cm above the liquid nitrogen, then were immersed in liquid nitrogen immediately after 10 minutes. Qualitative parameters of sperm included viability, total motility (TM), progressive motility (PM), plasma membrane integrity (HOST), percentage of abnormal sperm and acrosome membrane damaged was studied in days 0, 15 and 30 of freezing process.

Percentage of viability, total motility, progressive motility, plasma membrane integrity (HOST) of sperm decreased significantly (p < 0.01). Also the percentage of acrosome membrane damage and abnormal sperm increased during experiment (p < 0.01). However, the addition of different levels of Nano Selenium to semen extender significantly increased the percentage of viability, percentage of sperm with total motility, progressive motility and health plasma membrane (positive HOST) on days 0, 15 and 30 compared with control group (p < 0.01). Also, the addition of different levels of Nano Selenium to extender significantly decrease the percentage of abnormal sperm and acrosome membrane damaged (p < 0.01).

According to the results of this study, the addition of different levels of Nano Selenium, especially Nano Selenium 1 µg/ml to ram sperm extender increases the quality of sperm during storage.

The use of the plant Tribulus Terrestris can improve semen quality in some mammals and birds. However, little information is available about adult male lambs. The purpose of this experiment was to investigate the effect of different levels of Tribulus terrestris on quantitative and qualitative characteristics of sperm of mature male lambs. In this study, 18 lambs of the arabi lambes age 3 monthes and average body weight 16.14±2 were divided into three groups of six in a completely randomized design, and received different levels of Tribulus terrestris including three groups: 1) without Tribulus terrestris (control), 2) 15 g/kg DM Tribulus terrestris and 3) 30 g/kg DM Tribulus terrestris for 5 months. The spermatogenesis was carried out by the electrocautery device two times a week from lambs after feeding Tribulus Terrestris. Quantitative and qualitative semen parameters including seminal volume and pH, sperm concentration, motility, survival and morphological abnormalities of sperms were evaluated. Results showed, the lowest semen volume was in the control group (p < 0.05). The highest concentration, motility and survival of sperm were from 15 g of Tribulus terrestris (p < 0.05). The level of 30 g of Tribulus terrestris showed the highest morphological abnormalities of sperm (p < 0.05). Mean pH was not affected by experimental treatments (p < 0.05). In conclusion, considering to all the measured parameters, adding 15 g of Tribulus terrestris per kilogram dry matter to the diet improved the quantitative and qualitative characteristics of sperm in the freshly mature arabi lambs.

Some previous researches have shown that dietary supplementation with some vitamins and trace elements can increase the ability of animals semen preservation during liquid storage or freezing. Therefore, the aim of this study was to investigate the effect of dietary supplementation of organic selenium and chromium on ram semen quality during liquid storage at 4 °C.

Numbers of 16 Mehraban rams, with ages ranging from 2-4 years and 69.75±10.44 kg average body weight, were divided into four groups of four and were fed by a basal diet consist of 70% forage and 30% concentrate. The experimental design was 2×2×4 factorial experiment based on randomized complete block design. The experiment lasted for 60 days. In this experiment, three factors were including selenium at two levels (0 and 600 µg/day/ram selenium as yeast-selenium), chromium at 2 levels (0 and 1000 µg/day/ram chromium as chromium-methionin) and time at 4 levels (0, 24, 48 and 72 hours after storage). Chromium was fed to rams using edible capsule shells and also selenium by drencher after dissolving in distilled water. In the last three weeks of the experiment, semen samples were collected 3 times. Semen samples after collection was diluted with tirs-egg yolk base extender and stored at 4°C for 72 h. Sperm cells viability, membrane integrity, abnormal morphology, total motility and progressive motility and some motion parameters is detected by computer-assisted sperm analysis at 0, 24, 48 and 72 hours after storage.

The effects of chromium, time and their interaction and also selenium × chromium interaction, selenium × chromium × time interaction were significant for sperm viability and membrane integrity (p < 0.05). The effect of selenium and selenium × chromium interaction was not significant for viability (p>0.05) but, the effect of chromium, selenium and time and also their interaction for membrane integrity was significant (p < 0.05). The viability reduced over time significantly (p < 0.05) and sperm viability in rams receiving the diet contained 1000 μg chromium, was higher than diet without chromium after storage at 4°C for 48 and 72 hours (p < 0.05). In addition, sperm viability in rams receiving the diet contained 600 μg selenium was higher than diet without selenium after storage at 4°C for 48 hours (p < 0.05). Adding 1000 μg chromium to diet containing 600 μg selenium, increased significantly sperm viability after storage at 4°C for 48 and 72 hours (p < 0.05). The membrane integrity reduced over time significantly (p < 0.05) and sperm membrane integrity in rams receiving the diet contained 1000 μg chromium was higher than other groups after storage at 4°C for 24 and 48 hours (p < 0.05). The effect of selenium and chromium and also their interaction was not significant for computer detected parameters (p < 0.05). Interaction of selenium and chromium for abnormal morphology was significant (p < 0.05) but the effects of chromium, selenium, time and their interaction was not significant (p>0.05). Total motility, velocity in curvilinear line, velocity in straight line, velocity in average path decreased over time during semen storage at 4°C (p < 0.05) and diet supplementation of selenium and chromium did not change these parameters (p>0.05). Progressive motility, abnormal morphology and straightness did not affected by time, selenium and chromum.

Generally, dietary supplementation of organic selenium and chromium in rams leads to increased sperm viability and membrane integrity during chilled liquid storage. Dietary supplementation of selenium and chromium did not change computer detected parameters during semen storage at 4°C. In general, the use of organic selenium and chromium in the diet of rams may increase the ability of sperm storage for artificial insemination.

In this research the effects of different levels and particle sizes of zinc oxide on reproductive performance of hens and roosters of broiler breeders were evaluated. A total of 200 female Ross 308 broiler breeders in a completely randomized design with 4 treatments, 5 replicates and 5 birds per each replication, and 24 male Ross 308 broiler breeders in a completely randomized design with 4 treatments, 6 replicates and one bird per each replication at 54 weeks of age were used. The experimental treatments included the diets containing 70 mg zinc from zinc oxide with large particle size (LPZnO-70), 100 mg zinc from zinc oxide with large particle size (LPZnO-100), 70 mg zinc from zinc oxide with small particle size (SPZnO-70) and 100 mg zinc from zinc oxide with small particle size (SPZnO-100). The results indicated that the experimental treatments had no significant effect on egg production, body weight, egg weight and egg yolk weight. In the entire experimental period, the egg shell thickness was significantly higher in treatment containing SPZnO than LPZnO (P<0.001). The highest and lowest fertility and hatchability ratioes were observed in treatments containing SPZnO-100and LPZnO-70, respectively. The highest semen volume was observed in roosters of treatments containing SPZnO-100 and LPZnO-100 (P< 0.05). Generally, it can be concluded that SPZnO has a more desirable effect on reproductive parameters of broiler breeders due to its higher bioavailability. Since the level of 100 mg zinc per kg of diet from both sources of zinc oxide resulted in better reproductive performance, the utilization of SPZnO-100 might be recommended.

According to an increasing interest for using artificial insemination by sheep producers, finding methods to maintain frozen semen fertility would be important and applicable. One of the freezing semen disadvantages is increasing of free radicals led to enhanced sperm membrane peroxidation then consequently causes lowered semen fertility after thawing. Some plants bearing high levels of plant antioxidants can help to decrease semen peroxidation. Cysteine, in addition, is a precursor of glutathione peroxidase being the most important antioxidant property in semen. Therefore, the aim of the study was to investigate the effect of adding different levels of ethanol extract of Mentha Longifolia (0, 100 and 200 microL/ml) and cysteine (0, 5 and 10 mmol/l) into ram semen on fertility features after freezing and thawing processes.

In the study, semen samples were collected from four mature, fertile Baluchi rams (> 2y-old) by artificial vagina and after initial evaluation were pooled together. Then, samples were extended in an extender based on Tris-egg yolk. Treatments were arranged as a 3×3 factorial experiment including nine treatments and six straws were assigned to each as replicate. After treating, the straws were filled by the equal amount of semen and after chilling in refrigerator (4 °C) for two hours were frozen in liquid nitrogen container. After thawing the straws at day 10, sperm motility parameters were assayed by CASA. Furthermore, the percentages of viability, plasma membrane integrity and normal morphology of sperms were evaluated by microscope. Data were analyzed by SAS software using GLM procedure and the means comparison was performed by Tukey-Kramer test at 5% error.

Results of the study showed that the effects of adding horsemint extract and cysteine into the extender and the interaction between them on the all motility and viability parameters of frozen ram sperm after thawing were significant (p<0.05), exempt for sperm morphology where only the effect of adding cysteine was significant. Means comparison showed that the greatest sperm total motility, plasma membrane integrity, linearity and curvilinear velocity were observed in cysteine10 treatment (without horsemint extract), while the greatest sperm viability, straight line velocity and average path velocity were seen in cysteine5 treatment (without horsemint extract). The greatest sperm progressive motility percentage was observed in horsemint200 group (without cysteine).

In general, considering to the beneficial effect of adding cysteine into the extender based on tris- egg yolk on the most of fertility parameters of ram frozen semen after thawing, using levels of 5 and 10 mmol cysteine per ml extender in order to maintain fertility of frozen ram semen is recommended.

The effect of feeding L-carnitine during pre-puberty on the quality parameters of fresh and frozen-thawed semen by using 12 Ross broiler breeder males (12 weeks) for 18 weeks, in a completely randomized design with three treatments (0, 250 and 500 mg / kg of L-carnitine in the diet) and four replications was performed. From the age of 26 to 29 weeks, semen collection was performed using abdominal massage. The sperms taken each time after dilution (with Beltsville diluent) were divided into two parts, one section was frozen and the other part was immediately examined. Motility (total and forward), viability, morphology, membrane functionality and lipid peroxidation parameters were evaluated. In fresh sperm, the correlation between L-carnitine and abnormalities was negative linear, and with viability was positive linear (P<0.05). Quadratic analysis was significant in forward Motility and MDA concentration (P<0.05). Birds that use diets containing L-carnitine, In terms of forward motility, viability, morphology and MDA concentrations in fresh sperm, And these traits, with the total motility and integrity of the plasma membrane of frozen sperm, were higher in comparison to the control group (P<0.05). Also, in the sperm after frozen-thawed, the correlation between L-carnitine and Motility (total and forward), viability and membrane integrity were positive linear (P<0.05), and the correlation between L-carnitine and MDA concentration was negative linear (P<0.05). The correlation between L-carnitine and Motility (total and forward), membrane integrity and MDA concentration were quadratic (P<0.05). According to the results, Dietary L-carnitine supplementation in pre-puberty improves the qualitative traits of sperm before and after freezing in the breeder broilers.

This study was conducted to assess the effects of dietary flaxseed and sesame oils, on the semen parameters, fatty acid composition of sperm as well as the fertility and hatchability eggs from aged roosters. In a completely randomized design, 24 Ross-308 roosters (aged 45 week) assigned to four groups, comprising six replicates and one bird in each. The birds received different diets including basal diet (control), basal diet supplemented with 2% flaxseed oil (FO), basal diet supplemented with 2% sesame oil (SO) and basal diet supplemented with 1% flaxseed oil and 1% sesame oil (MO). The diets were iso-caloric and iso-nitrogenous, containing the same level of vitamin E. The roosters were fed diet for 60 days, during which semen samples were collected on 1st, 20th, 40th and 60th days and the samples were tested for different characteristics. The results indicated that different diets affected semen qualities, except semen volume and the morphology. The concentration, progressive motility, MDA as well as viability of sperms were significantly different during different times of the experiment. The sperm quality parameters including total and progressive motility as well as MDA turned out to improve in the roosters fed FO or MO. Furthermore, the integrity of sperm membrane, DHA and DPA concentration, as well as the fertility were higher in the treatment group containing FO. It seems that supplementation of aged rooster’s diet with flaxseed oil or mixed oils, together with vitamin E improves the semen qualities and it can be applied as an appropriate strategy to preserve the reproductive performance of aged rooters.

The increasing use of artificial insemination (AI) in the poultry industry emphasizes the need for the sharing of good quality sperm. The evaluation of semen quality characteristics of poultry gives an excellent sign of their reproductive potential and has been reported to be a major determinant of fertility and subsequent hatchability of eggs. In poultry industry in order to take advantage of new AI techniques, proper storage of poultry semen is needed. As poultry semen is viscous, highly concentrated, with low volume, containing 6 (chicken) to 12 (turkey) billion spermatozoa/ mL, the extension of good semen with a proper diluent is required prior to AI and storage. Reproduction performance is affected some factors include; nutrition, genetic, management, and environment. One of the fundamental aim in poultry industry is production of fertile eggs with high hatchability. In reproduction physiology, male and female are important, but sire effect is more than dam because of breeding program, semen samples could be applied for females. In layer industry, number of females are more than males; so, artificial insemination is an important tool for genetic diversity and high reproductive performance. One of the major problems in artificial insemination is low quality of poultry semen. Freezing and thawing have adverse effects on sperm viability. Since lipids (poly unsaturated fatty acids) are important part of plasma membrane of sperm cells, it could be destroyed in short times of storage in a liquid condition. It has been reported that antioxidants can be employed to prevent sperm damage during chilling storage condition (Malo et al. 2011). It is well known that sperm of farm animals is highly sensitive to oxidative stress (Surai et al. 1998). The high concentration of polyunsaturated fatty acids in the cell membrane of the sperm is also considered as another key factor for susceptibility to the oxidative stress. The process of semen production of reactive oxygen species (ROS) may lead to impairment of sperm morphology, motility, and finally viability. Pharmacological plants have some antioxidant components, which decrease free radical levels in organism. One of the major problems in sperm viability is sperm fragmentation. In this case, nucleus chromatin of sperm damages or splits. In some cases, the motility and normality of the semen analysis is acceptable, but the sperm nucleus of sample is damaged.  Different tests are available for detection of sperm DNA damages like COMET assay, TUNEL assay, SCSA, and Sperm Chromatin Dispersion (SCD) test. Among these assays, the SCD test is a simple and low-cost method for the analysis of sperm DNA fragmentation (Shanmugam et al., 2014). This method is based on the principle that sperm with DNA fragmentation fails to produce halo of dispersed DNA loops when mixed with agarose followed by acid denaturation and nuclear protein removal. The halos can be visualized using bright-field microscopy after staining with Wright’s stain. This study was designed to evaluate the effects of Rosemary extract on rooster semen parameters during chilling storage condition at 4ºC.

Dimension of individual cages were 85×70×70cm. Photoperiod program was applied based on 10 h darkness and 14h light. Water and feed were ad libitum. Rooster were fed corn and soybean meals based on 2107 kcal and 12% protein per Kg of dry matter. The semen was collected twice a week for six times from 10 mature roosters using abdominal rubbing method. As the rooster responded to abdominal rubbing by everting its cloaca, the ejaculate was collected using a graduated pipette; then, total volume was recorded. Urine or fecal contamination was discarded. Fresh semen was evaluated for sperm concentration using a haemocytometer. The semen samples were diluted by Sexton extender and evaluated at 25-27 °C. Then, the samples were divided into six equal parts. Different treatments were including: 0(control), 15, 25, 35, 45 and 60 µg/ml of Rosemary extract. Rosemary extract were added to each part and kept at 4oC for 72h. The parameters included motility, viability and plasma membrane integrity were evaluated at 0, 24, 48 and 72 h after storage. Also, DNA fragmentation was evaluated 48h after storage at 4 oC. In this method, sample is affected by acid treatment and denatures sperm chromatin. For elimination of proteins from chromatin, DNA strands are scattered around the sperm head and then, a halo forms around the sperm head.  With staining, halo is visible under the microscope. Halo in fragmented sperm is very small.   Data of the experiment were analyzed by Proc ANOVA SAS software (repeated measurement) and means were compared by Duncan's multiple range test.

Sperm during time of storage undergoes different changes and potentially undergoes mechanical and oxidative damages. At the time of storage in liquid condition, free radicals are released and sperm performance decreases significantly. The results of this experiment revealed that applying 35 µg/ml Rosemary extract could decrease DNA fragmentation (P<0.05). It seems that this concentration of Rosemary extract has antioxidant properties in rooster semen. Avian sperm have high concentration of long chain polyunsaturated fatty acids within the phospholipids and therefore, are prone to peroxidation damage (Blesbois et al. 1997). Also, after 48h of storage, motility, viability and sperm plasma membrane integrity were lower in treatment 60µg/ml Rosemary extract than control group (P<0.05). Finally, it was demonstrated that 48h after storage, motility, viability and sperm plasma membrane integrity in samples treated by 35µg/ml Rosemary extract were higher than control group (P<0.05). High level of reactive oxygen species (ROS) in oxidative stress is related to DNA fragmentation. Free radicals oxidize purine and pyrimidine bases of DNA structure of sperm and it causes damage DNA structure. The prevention of free radicals is the first defense line of antioxidants against oxidative damage. Rosemarry extract improved rooster sperm quality after thawing (Shafigh et al. 2016).  This is the first study to report the use of DNA fragmentation test in rooster sperm in Iran. This is a simple technique requiring no sophisticated instruments when Wright stain is used for visualization of the halos. According to the results of this study, it seems that applying 35µg/ml Rosemary extract can be useful in rooster semen at 4 oC. Rosemary extract effectively reduced DNA fragmentation. This effect seems to be associated with the high motility of treatment (35µg/ml Rosemary extract) compare with other treatments.