The effect of peroxisome proliferator-activated receptor gamma (PPARγ) as a mediator of dietary fatty acids and thiazolidinedione in pulmonary arterial hypertension induced by cold stress of broilers.

The objective of this experiment was to evaluate the effects of dietary fish oil and pioglitazone as peroxisome proliferators-activated receptor gamma (PPARγ) activating ligands on the reduction of cold-induced ascites in broiler chickens. A total of 480 one-day-old (Ross 308) male chicks were randomly allocated to four treatment groups with eight replicates of 15 birds each. The following treatments were used: 1) ambient temperature (negative control), with basal diet; 2) cold-induced ascites (positive control), with basal diet; 3) cold-induced ascites, with basal diet +10 mg/kg/day pioglitazone and 4) cold-induced ascites, with basal diet +1% of fish oil. When compared with the positive control, body weight gain was higher (P ≤ 0.05) for broilers fed diets containing fish oil and pioglitazone at 28, 42, and 0-42 d. Broilers under cold-induced ascites had the highest blood pressure at 21 and 42 d, while fish oil and pioglitazone treatment reduced the blood pressure (P ≤ 0.05). Red blood cells, white blood cells, hematocrit, erythrocyte osmotic fragility, bursa of Fabricius and spleen weights were improved (P ≤ 0.05) for chickens fed fish oil diets and pioglitazone compared to the cold-induced ascites (positive control). Exposure to cold temperature resulted in an increase in plasma T3 and T3/T4 ratio and decline in plasma T4 (P ≤ 0.05). In conclusion, PPARγ agonist pioglitazone and fish oil as source of omega-3 polyunsaturated fatty acid could be used as a strategy to reduce the negative effects of pulmonary arterial hypertension and ascites in broiler chickens.

Peroxisome proliferator-activated receptor gamma (PPARγ) activation: a potential treatment for ascites syndrome in broiler chickens.

Ascites (serous fluid accumulation in the abdominal cavity) has been observed worldwide in fast growing broilers. Pulmonary vascular remodeling is an important pathological feature of broiler ascites syndrome. Peroxisome proliferators-activated receptor gamma (PPARγ) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) are expressed in pulmonary vascular endothelial cells and vascular smooth muscle cells (VSMC) where they participate in the regulation of normal pulmonary vascular function. The objective of the present study was to investigate the effects of omega-3 fatty acids (found in fish oil) and pioglitazone (PIO) as natural and synthetic PPARγ ligands supplementation on PPARγ and PGC-1α expression in the prevention of pulmonary arterial hypertension (PAH) syndrome in broiler chickens. The experiment was conducted with 4 treatment groups: 1) negative control, normal temperature conditions with basal diet; 2) positive control, low-temperature conditions with basal diet; 3) positive control + 10 mg PIO/kg of weight/d and 4) positive control + 1% FO. Each treatment had 5 replicates. Ascites heart index (RV/TV) was significantly (P < 0.05) reduced in chickens receiving FO (0.20) and PIO (0.21) compared to the positive control group (0.26). The addition of PIO in broilers under cold-induced ascites significantly increased the expression of PPARγ (9.44) and PGC-1α (5.81) genes in lung tissue compared to the negative control group (1.03, P < 0.05). Proliferative indexes of VSMC in pulmonary arteries such as PMT, PIT, and percentage wall thickness were significantly elevated in positive control group, indicating that pulmonary vascular remodeling occurred following VSMC proliferation in ascites. The vessel internal diameter was increased in FO and PIO groups. Based on these results, activation and expression of PPARγ and PGC-1α genes as a critical regulator of pulmonary artery smooth muscle cell using ligands, especially PIO, can be effective in reducing the incidence of PAH in broiler chickens.

Determining the Optimal Dosage of Lecithin Nanoliposome in Rooster Semen Freezing Medium and Fertility Potential.

Introduction: Lecithin nanoliposome (nano-LPO), with its cryoprotective properties, is considered to enhance the performance of a traditional semen cryoprotectant. Objective: To determine the optimal dose of lecithin nano-LPO added to the rooster semen extender. Materials and Methods: Semen samples collected weekly from eight broiler breeder roosters were mixed and aliquoted into five equal subsamples, during the five successive weeks. The subsamples were then diluted with a semen extender containing 0%, 0.5%, 1%, 1.5%, or 2% of lecithin nano-LPO. Post-thawed semen quality attributes, including sperm motility and velocity parameters, plasma membrane functionality, mitochondrial membrane potential (MMP), apoptosis-like changes, and fertility potential, were evaluated. Results: Total motility and velocity parameters, including curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity µm/s (VAP), straightness (STR), linearity (LIN), lateral head displacement (ALH), and wobble (WOB) were quadratically (p < 0.01) influenced by graded levels of lecithin nano-LPO, such that the highest values were obtained when 1% of lecithin nano-LPO was used. Treatments had no significant effect on plasma membrane functionality; however, MMP (p < 0.08) and percentages of live and dead spermatozoa (p < 0.05) quadratically responded to increasing levels of lecithin nano-LPO, where the best outcome was found when about 1% of lecithin nano-LPO was used in the semen extender. The percentage of apoptotic spermatozoa cubically responded to increasing levels of lecithin nano-LPO (p ≤ 0.07). No significant trend of fertility rate was found in response to addition of lecithin nano-LPO levels. Conclusions: Supplementing an extender with 1.10% of lecithin nano-LPO is shown to be the optimal dose associated with the most improvement in post-thawed rooster sperm velocity measurements.

Sperm flow cytometric parameters, antioxidant status, and testicular histomorphology in roosters fed diets supplemented with camphor.

Genetic selection based on the high growth rate and consequently high slaughter weight in broiler chickens has caused many problems in broiler breeders. A negative correlation between growth and reproductive traits has declined semen quality and fertility in roosters. The present study aimed to evaluate the effects of camphor levels on some reproductive parameters included semen parameters, antioxidant status, and testicular development in broiler breeder roosters. Thirty-five ROSS 308 broiler breeder roosters were divided into 5 groups to receive camphor (C) levels: C0, C50, C250, C750, and C1000 ppm for 12 consecutive weeks (31-43 wk). Body weight, seminal volume, sperm concentration, and percentage of live and morphologically normal sperm were not affected by diets (P > 0.05), however, significantly were changed by bird age over the experiment (P < 0.05). Semen quality factor (SQF) significantly was affected by both diets and age (P < 0.05). Mitochondrial activity, apoptotic-like changes, and DNA fragmentation were improved in the groups fed camphor levels compared to the control group (P < 0.05). Testes weight (left, right, and combined weights) and gonadosomatic index were increased linearly by the camphor supplementation (P < 0.05). The serum activity of glutathione peroxidase (GPX) was not affected by treatments, however, superoxide dismutase (SOD) activity, ferric ion reducing antioxidant power (FRAP), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity were significantly higher in C50, C250, and C750, respectively (P < 0.05). The lower malondialdehyde (MDA) content in the testes and liver samples was observed in C750 (P < 0.05). Excluding the number of Sertoli cells and blood vessels, other histomorphological traits of testes showed one of the linear or quadratic responses to the camphor levels (P < 0.05). It can be concluded that camphor as an antioxidant source may improve reproduction performance in roosters.

Enhancement of sperm quality and fertility-related parameters in Hubbard grandparent rooster fed diets supplemented with soybean lecithin and vitamin E.

The purpose of this study was to investigate the effect of dietary supplementation of different levels of soybean lecithin and vitamin E on semen quality parameters and some reproductive hormones in Hubbard grandparent roosters. The experiment was conducted in a 3 × 2 factorial arrangement with 3 levels of soybean lecithin (0, 1, and 2%) and 2 levels of vitamin E (0 and 300 mg/kg). Semen samples were collected on d 0, 20, 40 and 60 of the experiment and analyzed. Adding 1% soybean lecithin and vitamin E into the diet increased semen volume and sperm concentration, membrane integrity and viability (P < 0.05). Supplementing diets with 1 or 2% lecithin in addition to vitamin E significantly improved total motility and progressive motility (P < 0.05). Vitamin E significantly increased the amplitude of lateral head displacement (ALH) of sperm (P < 0.05). Although there was no effect on LH and FSH when diets were supplemented with vitamin E and 1 or 2% lecithin, testosterone concentration was increased (P < 0.05). Malondialdehyde (MDA) concentration was significantly lower in all 3 treatments containing vitamin E (P < 0.05). It can be concluded that supplementation of rooster diets with vitamin E and 1% lecithin can improve fertility related parameters in Hubbard grandparent roosters.

Supplementation of freezing medium with encapsulated or free glutathione during cryopreservation of bull sperm.

The objective was to compare effects of encapsulated or free glutathione (GSH) on the quality of frozen-thawed bull sperm. Ejaculates were collected via artificial vagina from six mature Holstein bulls once weekly for 6 weeks. All ejaculates had motility ≥70%, sperm concentration ≥1.0 × 109 /ml and ≤15% morphologically abnormal sperm. Each week, semen was pooled and diluted with lecithin-based extenders containing various concentrations of encapsulated (E0, E1, E2.5 and E5 mM) or free (F0, F1, F2.5 and F5 mM) GSH, with total glutathione content determined before and after cryopreservation. Total GSH in fresh semen was (mean+SEM) 4.8 ± 0.2 nmol/108 sperm, whereas in frozen-thawed semen of group F0 (control), it decreased to 1.4 ± 0.2 nmol/108 sperm, a 70.8% reduction (p < .05). In addition, total GSH in frozen-thawed semen from groups E2.5, E5 and F5 were 2.4 ± 0.2, 2.8 ± 0.2 and 1.8 ± 0.2 nmol/108 sperm, respectively (E5 versus. F0, p < .05). Compared to group F0, frozen-thawed sperm from group E2.5 had greater (p < .05) percentages of sperm that were viable (Annexin-V) (61.1 ± 1.8 versus. 71.1 ± 1.8) and that had cell membrane integrity (eosin-nigrosin) (64.5 ± 3.1 versus. 80.0 ± 3.1). Furthermore, frozen-thawed sperm from group E2.5 had the numerically highest total and progressive motility (CASA) and cell membrane functionality (HOS) and the lowest percentage of early apoptotic sperm (Annexin-V). However, acrosome membrane integrity (PSA) of E5 had the lowest mean (p < .05), whereas E2.5 caused a small nonsignificant decrease (69.1 ± 1.4%) compared to E0 and F0. In conclusion, 2.5 mM encapsulated GSH in semen extender significantly improved the quality of frozen-thawed bull sperm.

Effects of dietary L-carnitine on puberty indices in the young breeder rooster.

The aim of current study was to investigate the effect of dietary L-Carnitine (LC) in immature roosters on reproductive hormones, lipid profile and testicular histology at the time of maturity. Eighteen 12-wk-old breeder roosters (Ross 308) of similar weights were randomly allocated into 3 dietary treatments (LC-0: basic diet, LC-250: basic diet + 250 mg LC/kg of diet, LC-500: basic diet + 500 mg of LC/kg of diet) in 6 replicates. The feeding program and photoperiod regimen were performed based on ROSS 308 management handbook. Dietary LC supplementation markedly improved testicle weight and testicle index (p < 0.05). Comb height was also affected by LC supplementation (p < 0.05). The testicle weight and index, comb height, and shank lengths improved linearly with increasing levels of dietary LC (p < 0.05). The LC-250 and LC-500 diets significantly improved the number of sertoli cells (NSC), height epithelium seminiferous tubules (HEST), seminiferous tubules diameter (STD), spermiogenesis index (SI) and tubular differentiation index (TDI) of rooster's testis tissue (p < 0.05). The number of seminiferous tubules (NST) was affected by of the amount of LC (p < 0.05). The roosters on the LC-250 mg/kg diet had longer HEST compared to roosters that received the LC-500 mg/kg diet (p < 0.05). Testicular histology parameters increased in a linear and quadratic manner in response to increasing levels of LC (p < 0.05). Dietary LC significantly increased (p < 0.05) plasma concentrations of testosterone, GnRH, LH, FSH and High-Density Lipoprotein (HDL), but reduced the plasma concentration of Low-Density Lipoprotein (LDL). However, no significant differences were observed between LC-250 and LC-500 groups in these parameters. Plasma testosterone, GnRH, LH, LDL and HDL were affected in a linear and quadratic manner in response to increasing levels of LC (p < 0.05). Similarly, FSH increased linearly with increasing dietary LC (p < 0.05). Thus, adding up to 250g of LC per kg of the rooster chicken can improve reproductive hormones, blood lipids and testicular histology parameters at the time of maturity.

Improving seminal quality and reproductive performance in male broiler breeder by supplementation of camphor.

The current study was conducted to evaluate the effect of dietary camphor levels as a medicinal feed additive to improve semen quality, antioxidant capacity, reproductive hormones, and reproduction performance in roosters. For this purpose, thirty-five 29-wk-old Ross 308 broiler breeder roosters randomly were assigned to five experimental groups (seven birds/group) and received five doses of camphor containing 0, 50, 250, 750, and 1000 mg camphor/kg of feed for 12 wk consecutive. Semen quality parameters and motion characteristics of sperm were evaluated every 28 days and semen antioxidant capacity and plasma reproductive hormones concentration were tested at the end of the experiment. Also, at the end of the experiment, reproductive performance was assessed using artificial insemination. Among seminal quality parameters, sperm forward motility (88.96 vs 82.56%) and percentage of abnormal sperm (14.75 vs 15.86%) were improved in roosters fed 50 mg camphor/kg of feed compared to the control group (P < 0.05). Overall percentage of live sperm and plasma membrane integrity exhibited the quadratic responses to the levels of camphor (P < 0.08). The motion characteristics of sperm including progressive motility (28.81 vs 21.77%), average path velocity (VAP, 33.35 vs 26.83 µm/s), progressive velocity (VSL, 19.78 vs 16.48 µm/s), curvilinear line velocity (VCL, 52.87 vs 44.38 µm/s), the amplitude of lateral head displacement (ALH, 2.92 vs 2.46 µm) were improved in roosters fed 50 mg camphor/kg of feed compared to the control group (P < 0.05). However, dietary camphor levels linearly increased the percentage of linearity (LIN) and straightness (STR) (P < 0.05). A significant decrease in seminal plasma concentration of malondialdehyde (MDA) and an increase in superoxide dismutase (SOD) activity were observed in birds fed 1000 mg camphor/kg of feed (P < 0.05). Testosterone concentration was considerably increased by doses of 50 and 250 mg camphor/kg of feed compared to control (4.68, 4.79 vs 3.88 ng/mL) (P < 0.05). FSH and LH concentrations were not affected by camphor supplementation (P > 0.05). In the artificial insemination, fertility rate from both 50 mg camphor/kg of feed (88%) and 250 mg camphor/kg of feed (84%) was higher than control (75%) (P < 0.05). In conclusion, low levels of camphor, especially 50 mg camphor/kg of feed, improved seminal characteristics and, reproductive performance of roosters. Further researches are needed on the effect of higher levels of camphor and divulge of underlying mechanism on male's reproductive function.

Effects of dietary alpha-lipoic acid supplementation on the seminal parameters and fertility potential in aging broiler breeder roosters.

High levels of polyunsaturated fatty acids in avian sperm cause more susceptibility to lipid peroxidation. Aging in roosters reduces the antioxidant capacity of sperm and thus fertility. The purpose of this study was to investigate the effects of different levels of alpha-lipoic acid (ALA) as a feed supplement to improve the semen quality and fertility parameters of aged broiler breeder roosters and identification of its most effective level. A total of forty-two roosters at 45 wk of age were randomly assigned to 7 treatments (0, 15, 40, 70, 95, 120, and 145 mg ALA/bird per day) for 8 wk. Semen parameters and body weight were assessed biweekly, and testosterone plasma levels were determined in the 8th wk of the experimental period. Artificial insemination was performed at the end of the experiment to evaluate the fertility potential. The dietary administration of ALA had no significant effects on body weight, semen volume, average path velocity, linearity, straightness, wobble, the amplitude of lateral head displacement, beat-cross frequency, sperm concentration, morphology, plasma testosterone level, fertility, or hatchability (P > 0.05). Alpha-lipoic acid supplementations resulted in a significant decrease in seminal malondialdehyde concentration and immotile (type D) sperms (P < 0.05). The total motility, progressive motility (types A + type B sperms), curvilinear velocity, straight-line velocity, viability, and membrane integrity of sperm improved with ALA dietary supplementations (P < 0.05). With increasing ALA levels, improvement in semen parameters had an incremental trend until the level of 95 mg ALA. Thus, 95 mg dietary ALA as an antioxidant supplement can improve semen quality of aging breeder roosters while higher doses resulted in no further improvement.

Manipulation of fatty acid profiles in roosters' testes, alteration in sexual hormones, improvements in testicular histology characteristics and elevation sperm quality factor by L-carnitine.

The present study was designed to investigate the effects of different levels of l-carnitine (LC) on sperm quality factor (SQF), alterations in testis fatty acid profiles, testicular histology and reproductive hormones in young roosters. Eighteen broiler breeders (Ross 308) weighed at 3 months of age. They were randomly classified while each group had six birds. There were three experimental groups based on the LC concentrations (i.e. LC-0, LC-250, LC-500 mg per kg of diet). After two weeks of adaptation, semen samples were collected and evaluated for seminal attributes every two weeks (from week 24 to week 34). At the end of the experiments, four roosters from each treatment group were sacrificed in order to analyze testicular histology, testis fatty acid profiles and reproductive hormones. Supplementing the diet with two of the LC levels for 22 weeks caused significant rise in sperm concentration, viability and SQF compared to that of the control group (P < 0.05). Quadratic analysis in terms of number of seminiferous tubules and spermatogenesis index were significant (P<0.05). Tubular differentiation index improved linearly by the increasing levels of LC supplementation (P<0.01). The analysis of fatty acid profiles showed that LC significantly (P < 0.05) reduced the percentages of C14:0, C21:0, total saturated fatty acids, total odd-chain fatty acids and n-6/n-3 ratio. Moreover, LC significantly increased the percentage of C20:5n-3 (Eicosapentaenoic acid; EPA) (P < 0.05). Analysis of the correlation coefficient revealed that the SQF is in consistency with EPA (r = 0.98; P < 0.04). In contrast, SQF negatively and significantly correlates with odd-chain fatty acids (r = - 0.99; P < 0.001). The desaturation index for C16 fatty acids (16:1cis/C16:0) negligibly increased linearly as LC was added to the diet (P < 0.05). Furthermore, LC caused the roosters to have significant (P < 0.05) high levels of total testosterone and FSH concentrations. The concentration of LH in different treatment groups, however, turned out to be similar in response to the different levels of LC. In conclusion, long-term supplementation of rooster diet with LC can have beneficial effects on SQF and testis histology. The benefits include alterations in testicular histology, reproductive hormones and testicular fatty acid profiles.

Dietary L-carnitine affects the expression of genes involved in apoptosis and fatty acid metabolism in rooster testes.

Thirty-six 12-week-old breeder roosters (Ross 308) were randomly allocated into three groups to receive L-carnitine (LC): LC-0, LC-250 or LC-500 mg/kg of diet to evaluate the effects of dietary LC on the expression of apoptotic-related genes and desaturases and elongase mRNA transcript levels, in the cockerel testicles. Alteration of Bak (Bcl2 antagonist/killer), Bcl2, Cas3, Cas8, Cas9, Elovl2, Elovl4, Elovl5, Fads1, Fads2 and Scd expression at 24 and 34 weeks of age was compared by real-time quantitative PCR. The expression of Bcl2 and Elovl5 was significantly up-regulated (p < .05), while Cas8 expression (p < .05) and Bak/Bcl2 ratio were reduced (p < .02) in the cockerel testicles at 24 weeks of age. Although Bak mRNA abundance decreased by dietary LC, Bak/Bcl2 ratio was not affected by the treatments at 34 weeks of age. The expression of Cas3 was down-regulated, while Fads2 was up-regulated in the cockerel testicles by dietary LC at 34 weeks of age (p < .05). The results demonstrate the beneficial effects of LC supplementation in suppression of the Bak/Bcl2 ratio by altering Bak and Bcl2 mRNA abundance and, ultimately, prevention of apoptosis. Furthermore, LC increased the expression of Elovl5 and Fads2 genes which are involved in the metabolism of long chain fatty acids.

Peroxisome proliferator-activated receptors (PPARs) as a mediator of dietary fatty acids affects reproductive performance in broiler breeder roosters.

This study analyzed the effects of dietary sources of omega-3 and omega-6 fatty acids on semen parameters and fertility potential in broiler breeder roosters. The mRNA and protein profiles of peroxisome proliferator-activated receptors-γ (PPAR-γ) expression in sperm as potential mediator of FAs were considered. Roosters were categorized into three groups and received their diets for 24 weeks as follows: 1) control diet received a basal diet (CTRL); 2) Fish oil based diet (FO) received the basal diet supplemented with 15 g/kg of diet fish oil; and 3) sunflower oil based diet (SO) received the basal diet supplemented with 15 g/kg of diet sunflower oil. While the different diets had significant effects on semen parameters, the effect of sampling time was not significant. The effect of the diets on sperm parameters were significantly higher in the SO and FO groups in total motility, progressive motility, amplitude of lateral head displacement, linearity, straightness, wobble and viability (P ≤ 0.05). Fertility rate was significantly improved in the FO and SO groups (P = 0001). The highest value for PPAR-γ mRNA was observed in the SO group compared to other groups (P ≤ 0.05). Moreover, supplementation of the roosters' diets with FO and SO increased PPAR-γ protein expression compared to the control. It seems that PPAR-γ could be a strong potential mediator of the underlying mechanism of improvement in semen parameters and reproductive performance of roosters under the effects of both dietary omega-3 and omega-6 polyunsaturated fatty acids.

Evaluation of used Purslane extracts in Tris extenders on cryopreserved goat sperm.

This study aimed to evaluate the comparative effects of Purslane aqueous extract (PAE), Purslane methanolic extract (PME) and Purslane ethanolic extract (PEE on the quality of frozen-thawed goat spermatozoa. Collected semen with motility >75% and sperm concentration >1.0 × 109 sperm/ml was pooled and divided into 10 equal aliquots and supplemented by basic extender containing 25, 50 or 100 µg/ml of Purslane aqueous extract (PAE25µg/ml, PAE50µg/ml, PAE100µg/ml, respectively), basic extender containing 25, 50 or 100 µg/ml of Purslane methanolic extract (PME25µg/ml, PME50µg/ml, PME100µg/ml, respectively), basic extender containing 25, 50 or 100 µg/ml of Purslane ethanolic extract (PEE25µg/ml, PEE50µg/ml, PEE100µg/ml, respectively). Control diluent contained no additives. For the determination of sperm quality, frozen straws were thawed and then the sperm characteristics were assessed. Results indicated that higher (P < 0.05) percentages of total motility, viability, mitochondrial activity and lower percentages of malondialdehyde (MDA) for PAE50µg/ml, PME50µg/ml and PEE50µg/ml than those of the control. In addition, PME50µg/ml resulted in the highest) P < 0.05) total motility and the lowest (P < 0.05) MDA levels compared to other treatments. Compared to the control group, PME50µg/ml resulted in higher integrity (P < 0.05) of plasma membranes and in lower amounts of apoptotic and dead spermatozoa. PME50µg/ml and PAE50µg/ml showed higher (P < 0.05) percentages of progressive motility, DNA integrity and live post-thawed spermatozoa than those of the control. No significant differences in the motility, viability, mitochondrial activity and number of live sperms were observed between PME50µg/ml and PAE50µg/ml treatments. In conclusion, the results of this study indicated that 50 µg/ml purslane extracts could be used for the cryopreservation. However, the results of methanolic extract was more beneficial compared to other extracts.

Amelioration effects of n-3, n-6 sources of fatty acids and rosemary leaves powder on the semen parameters, reproductive hormones, and fatty acid analysis of sperm in aged Ross broiler breeder roosters.

This study was conducted to evaluate the effects of dietary polyunsaturated fatty acids (PUFA) sources and rosemary leaves powder (RLP) on the semen quality, fatty acid analysis, and some reproductive hormones of senescent broiler breeder roosters. Thirty-five 45-wk-old Ross breeder roosters were randomly divided into 7 groups (5 birds/group), and received following treatments including control group (basal diet), fish oil (2%), corn oil (2%), an equal (50:50%) proportion of fish oil and corn oil (50:50%), fish oil (2%) with 5 g/kg capsulated RLP, corn oil (2%) with 5 g/kg capsulated RLP, and an equal (50:50) proportion of fish oil and corn oil (50:50%) with 5 g/kg capsulated RLP of diet for 60 D, during which time their seminal characteristics were evaluated every 20 D. At the end of the trial (on day 60), semen samples were tested for determination of sperm fatty acid analysis, lipid peroxidation, and some reproductive hormones. Results showed that feeding fish oil and fish/corn oil with RLP was associated with an increase in docosahexaenoic acid (C22:6n-3) and docosatetraenoic acid (C22:4n-6) in sperm. The fish oil diet increased the proportion of n-3 fatty acids in sperm, and as a consequence, the (n-6)/(n-3) ratio also decreased (P < 0.05). RLP (5 g/kg) to the fish and fish/corn-oil (50:50%)-based diet resulted in improvement in sperm concentration, total motility (%), sperm progressive motility (%), membrane integrity, and viability in terms 0 to 60 day trial (P < 0.05). Diets and age interacted to positively affect sperm concentration and sperm membrane integrity. Also this herbal antioxidant decreased the seminal content of malondialdehyde (MDA) significantly (P < 0.05). Testosterone and LH serum levels of reproductive hormones were significantly higher in fish and fish/corn-oil with RPL (50:50%)-based diet than other groups (P < 0.05). It can be concluded that RLP as an antioxidant could remarkably improve the effects of n-3 and n-3/n-6 PUFA on sperm characteristics, seminal MDA, and hormones levels in aged breeder roosters. The susceptibility of semen to lipid peroxidation was increased in chickens fed without RLP. Future studies are needed to disclose the causal mechanisms involved.

Inclusion of ovine enriched serum with vitamin E and polyunsaturated fatty acids in the freezing medium: a new strategy to improve human frozen-thawed sperm parameters.

The objective was to evaluate the effect of inclusion of 2.5% and 5% ovine serum, enriched with vitamin E (Vit E) and fish oil (FO), in human sperm freezing medium. Serum samples were prepared from sixteen rams (n = 4) feeding on a without supplemented diet, and diets supplemented with Vit E, FO and Vit E + FO. Semen samples, from 60 normozoospermic men, were frozen in: (I) a commercial freezing medium (SpermFreeze™; control medium), (II) the commercial freezing medium containing foetal bovine serum, (III) the commercial freezing medium + nonenriched serum (serum group), (IV) the commercial freezing medium + Vit E enriched serum (Vit E group), (V) the commercial freezing medium + FO enriched serum (FO group) and (VI) the commercial freezing medium + Vit E + FO enriched serum (Vit E + FO group). Sperm total and progressive motility, morphology, viability and plasma membrane integrity were significantly higher (p ≤ .05) in Vit E and Vit E + FO groups compared with the control group. Mitochondrial membrane potential did not differ between treatments (p > .05). It was concluded that ovine serum enriched with vitamin E and vitamin E + FO improved the quality of human spermatozoa but enriched serum containing FO could not improve the sperm cryo-injuries.

Comparison of two different antioxidants in a nano lecithin-based extender for bull sperm cryopreservation.

The objective of the present study was to assess the effect of two different antioxidants, enzymatic compared with non-enzymatic, in a nano lecithin-based extender on post-thaw bull sperm quality. Semen samples (n = 36) were collected from six bulls. In the first experiment, 11 different extenders were prepared by adding five quantities of vitamin E (α-tocopherol) as a non-enzymatic antioxidant (VE: 0.1, 0.2, 0.4, 0.6 and 1.0 mM), or four quantities of glutathione peroxidase (GPx) as an enzymatic antioxidant (GPx: 0.5, 1, 2 and 3 mM) to the extender. Other extenders were a Control 1 (C1: Extender with ethanol) and Control 2 (C2: Extender without ethanol). Sperm motility (CASA), plasma membrane functionality test (HOST) and lipid peroxidation (MDA) were assessed to determine the optimal treatment in the first experiment. In the second experiment, the optimally supplemented group from the first experiment (GPx-1) was compared to C2 group. Apoptotic-like changes (Annexin staining), mitochondrial activity (Rhodamine-123 staining), acrosome integrity (PSA staining), DNA fragmentation (SCSA test) and in vitro embryo production capacity were evaluated. In the first experiment, there were the greatest percentages of plasma membrane functionality and least MDA (P ≤ 0.05) in sperm diluted GPx-1 group. In the second experiment, percentage of live sperm, blastocyst formation and hatching rate were greater (P ≤ 0.05) in the GPx-1 group compared with C2 group. In conclusion, data indicate adding 1.0 mM GPx as an enzymatic antioxidant to the nano lecithin-based extender can improve post-thaw quality and in vitro fertility of bull sperm.

Effects of CoQ10 on the quality of ram sperm during cryopreservation in plant and animal based extenders.

The purpose of this study was to evaluate the effect of different concentrations of CoQ10 in soybean lecithin (SL) or egg yolk (EY) extenders on ram semen cryopreservation. Semen samples were collected from five rams, twice a week, then diluted in the extenders (SL and EY) containing different concentrations of CoQ10 as follows: extender containing SL: 0 µM (control, SL/Q0), 1 µM (SL/Q1), 2 µM (SL/Q2), 5 µM (SL/Q5) and 10 µM (SL/Q10) CoQ10; extender containing EY: 0 µM (control, EY/Q0), 1 µM (EY/Q1), 2 µM (EY/Q2), 5 µM (EY/Q5) and 10 µM (EY/Q10) CoQ10. Sperm motion characteristics, membrane integrity, abnormal morphology, viability, apoptotic-like changes, mitochondria active potential, acrosome integrity and lipid peroxidation were evaluated after freeze-thaw process. The SL/Q1, SL/Q2, EY/Q1 and EY/Q2 resulted in greater (P ≤ 0.05) sperm total motility, progressive motility, membrane integrity and mitochondria active potential compared to the other groups. Acrosome integrity in the SL/Q0, SL/Q1, SL/Q2, EY/Q0, EY/Q1 and EY/Q2 groups was greater (P ≤ 0.05) than in the SL/Q5, SL/Q10, EY/Q5 and EY/Q10 groups. The SL/Q2 and EY/Q2 treatment groups had greater (P ≤ 0.05) sperm viability rates and less apoptotic-like changes and lipid peroxidation. The CoQ10 compound could be explored as a novel potential antioxidant for cryopreservation of ram semen because with used of this compound in the present study there was an improved post-thawed sperm quality.

Dietary supplementation with flaxseed oil as source of Omega-3 fatty acids improves seminal quality and reproductive performance in aged broiler breeder roosters.

Reproductive performance in aged broiler breeder roosters is not desirable. The purpose of this study was to assess the effects of dietary flaxseed oil and vitamin E on the semen parameters, hormonal profiles, fatty acid analysis of sperm, and fertility performance of aged roosters. Twenty four roosters were assigned into four treatments and received their diets as following: 1) basal diet as control group (CTRL), 2) basal diet supplemented with 200 mg/kg vitamin E (CTRL + VITE), 3) basal diet supplemented with 2% flaxseed oil (FLAX) and 4) basal diet supplemented with 2% flaxseed oil plus 200 mg/kg vitamin E (FLAX + VITE). Roosters were fed their diets for 60 days and then, different characteristics of reproduction in the roosters were examined during experiment. Different diets affected semen parameters (P ≤ 0.05) except semen volume and morphology. Various characteristics of semen were significant (P ≤ 0.05) during different times of experiment excluding the total motility, membrane integrity and morphology. The higher percentage of sperm concentration, total motility, progressive motility, viability and membrane integrity accorded with a lower lipid peroxidation were observed in the roosters fed diet of FLAX + VITE (P ≤ 0.05). A higher concentration of testosterone was detected in roosters fed FALX + VITE at day 60 (P ≤ 0.05). Moreover, the percentage docosapantaenoic acid (DPA), docosahexaenoic acid (DHA) of sperm were increased in the diet of FLAX (P ≤ 0.05). Finally, rate of fertility after artificial insemination was significantly higher in the diet of FLAX + VITE. It seems that supplementation of aged roosters' diet with flaxseed oil and VITE improves the semen performance and fertility potential that can be a suitable strategy to preserve the reproductive performance of aged rooters.

Supplementation of extender with coenzyme Q10 improves the function and fertility potential of rooster spermatozoa after cryopreservation.

The effects of coenzyme Q10 (CoQ10) has not yet been assessed for cryopreservation of rooster semen. The aim of this study was to evaluate the effect of different concentrations of CoQ10 in Lake extender for cryopreservation of rooster semen. The viability and apoptosis status, DNA fragmentation, abnormal morphology, motion parameters, membrane functionality, mitochondrial activity, acrosome integrity, lipid peroxidation, and fertility potential were evaluated after the freeze-thaw process. Semen samples were collected from ten roosters, twice a week, and then diluted in extender contained different concentrations of CoQ10 as follows: Lake without CoQ10 (control, Q 0), Lake containing 1 µM (Q 1), 2 µM (Q 2), 5 µM (Q 5), and 10 µM (Q 10) CoQ10. Supplementation of Lake with 1 and 2 µM CoQ10 resulted in greater sperm viability, total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, and fertility rate. Furthermore, the extent of lipid peroxidation in thawed spermatozoa treated with 1 and 2 µM CoQ10 was less than with the other groups. Different concentrations of CoQ10 had no effect on DNA fragmentation and sperm morphology. Results of the present study indicate that supplementation of Lake extender with 1 and 2 µM CoQ10 enhances the quality of rooster sperm after the freeze-thaw process.

Dietary fish oil supplemented with vitamin E improves quality indicators of rooster cold-stored semen through reducing lipid peroxidation.

Cockerel semen is sensitive to cooling, which limits chilled storage of semen for more than 24 h. Results of artificial insemination with cold-stored semen are not desirable. This study was conducted to evaluate the effects of dietary fish oil and vitamin E (vitE) for cold-storage of rooster semen and its effects on parameters of semen during 48 h cooling preservation. Roosters were assigned into four dietary treatments; 1) control group received a basal diet, 2) vitE group received a basal diet supplemented with 200 mg/kg vitE, 3) fish oil group (FO) received a basal diet supplemented with 2% fish oil and 4) fish oil and vitE group received a basal diet supplemented with 2% fish oil and 200 mg/kg vitE (FO + vitE). Semen samples were collected after 40 days of feeding and then diluted and cooled to 5 °C for preservation up to 2 days. Several quality indicators of sperm such as motion characteristics, membrane integrity, and viability, and abnormal morphology, activity of mitochondria, lipid peroxidation and acrosome integrity of the sperm were assessed at different times of storage (0, 24 and 48 h). None of sperm were significantly affected by the diets at the start of storage (0 h, p > 0.05). FO and FO + vitE improved the percentage of total motility, viability, and mitochondria activity at 24 h (P ≤ 0.05). After 48 h, only FO + vitE group produced the higher percentage of total motility, viability and membrane integrity (P ≤ 0.05). Lipid peroxidation was significantly reduced in sperm obtained from roosters fed diets of FO + vitE and vitE compared to FO and control (P ≤ 0.05) at times of 24 and 48 h. There was no significant difference between control and vitE groups in none of parameters (P > 0.05). Integrity of acrosome and abnormal morphology were not significantly affected by the diets (P > 0.05). Supplementation of roosters' diet with 2% fish oil and 200 mg/kg vitamin E improved the quality of cold-stored semen by supporting several indicators of sperm quality through reducing lipid peroxidation.