Egg yolk plasma enriched with ß-carotene through the diet of laying hens and adding it to the extender improves the quality of frozen semen in Arabic stallions.

Equine semen cryopreservation is one of the major procedures for the genetic conservation of rare and endangered genotypes. The current study was conducted to evaluate the effect of egg yolk plasma (EYP) enriched with ß-carotene as an antioxidant supplement on INRA-96 extender regarding freezing Arabic stallion sperm. For this purpose, ß-carotene various concentrations were utilized as a supplementary ingredient in formulating the diets of laying hens. Birds were randomly divided into four groups, fed with 0 (control), 500, 1000 and 2000 mg/kg in a supplemented diet with ß-carotene. Subsequently, various variants of enriched extender (INRA-96 + 2.5% glycerol [G]) were gained by adding 2% EYP from four treatment groups. The sperm characteristics, including motility, viability, morphology, plasma membrane integrity (HOS test), lipid peroxidation (MDA) and DNA fragmentation, were evaluated after thawing. According to the results obtained in this study, the addition of EYP from T2 and T4 (500 and 2000 mg/kg of ß-carotene in hens' diet) to the extender (INRA-96 + 2.5% G) leads to an increase in total motility (50.50% and 49.49%, respectively), progressive motility (32.6% and 31.8%, respectively), viability (68.7% and 66.1%, respectively) and plasma membrane integrity (57.7% and 50.6%, respectively). Moreover, lipid peroxidation (1.3 and 1.4 nmol/mL, respectively) and DNA fragmentation (8.6% and 9.9%, respectively) were diminished using the mentioned treatments. However, sperm morphology was not affected by the treatments. In the current study, we concluded that the optimal concentration of ß-carotene in the laying hen's diet (500 mg/kg) could reveal the best results about sperm quality. So, EYP enriched with ß-carotene acts as a valuable natural and safe supplementary material that could be exploited for enhancing stallion sperm quality in cryopreservation conditions.

L-carnitine improves quality parameters and epigenetic patterns of buck's frozen-thawed semen.

Buck sperm cryopreservation is an effective method to distribute qualified sperm for reproductive purposes, but this procedure reduces sperm quality. The current study was conducted to investigate the effect of L-carnitine (LC) on the quality and epigenetic patterns of buck's post-thawed semen. Semen samples were collected from five male goats twice a week and diluted in extenders supplemented with 0 (LC-0), 1 (LC-1), 5 (LC-5) and 10 (LC-10) mM LC. Samples were cryopreserved according to standard protocol. After thawing, motility characteristics, lipid peroxidation, membrane functionality, abnormal morphology, mitochondrial activity, acrosome integrity, epigenetic modifications, viability, apoptotic-like changes and DNA fragmentation were assessed. Samples supplemented with 5 mM LC showed greater (P ≤ 0.05) total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, DNA methylation, viability, and lower (P ≤ 0.05) apoptotic-like changes. Lipid peroxidation was lower (P ≤ 0.05) in LC-5 and LC-10 compared to the control group. Addition of LC to the cryopreservation extender had no effect (P > 0.05) on velocity parameters, abnormal morphology, histone modifications, or DNA fragmentation. In conclusion, supplementing the cryopreservation extender with 5 mM LC significantly preserves the quality of buck sperm after the cryopreservation process.

Influence of Eimeria spp. infection and dietary inclusion of arginine on intestine histological parameters, serum amino acid profile and ileal amino acids digestibility in broiler chicks.

Coccidiosis is considered to be one of the most important challenge in the poultry industry causes economic losses due to the destruction in the digestive tract of chicken. It disturbs amino acids profile and their digestibility, leading to weight lost and economic burden. Using dietary arginine may decrease the adverse effects of coccidiosis on chicken digestive tract. This study aimed to evaluate the effects of dietary inclusion of arginine on intestine histological parameters, serum amino acid concentration and ileal amino acid digestibility of broiler chicks infected with coccidiosis. A total number of 384 one-d-old broiler chicks (Ross 308) of mixed sex with initial weight of 42 ±â€¯2 g was allocated into 8 groups with 8 birds/pen from grower period. At 21 days of age, broiler chicks were infected with a mixture of Eimeria spp. Broiler chicks were divided into infected and un-infected groups and received arginine at recommended levels of 85, 100, 125 and 150 %. Intestinal morphology and lesions, serum amino acid concentration and ileal amino acid digestibility were evaluated. Broiler chicks infected with Eimeria spp. showed lower villus height and villus height: crypt depth ratio and also higher intestinal lesions (P < 0.05). Coccidia infection decreased the ileal amino acid digestibility for all studied amino acids and also reduced serum concentrations of amino acids, except lysine and isoleucine (P < 0.05). Dietary supplementation of arginine especially in higher levels significantly increased villus height and villus height:crypt depth ratio and decreased lesions (P < 0.05). Moreover, dietary supplementing of arginine increased the serum concentration of arginine (P < 0.05), but it did not have any significant effect on its digestibility (P > 0.05). In sum, coccidiosis decreases amino acid digestibility and serum amino acid concentration, but dietary inclusion of higher levels of arginine significantly improved histological parameters of broiler chicks infected with coccidiosis.

Effect of egg yolk plasma and soybean lecithin on rooster frozen-thawed sperm quality and fertility.

This experiment was conducted to study the effects of egg yolk plasma (10%, 15% and 20%), soybean lecithin (0.5%, 1% and 1.5%) and whole egg yolk (WEY) (control) on post-thawed sperm quality, hatchability and fertility outcomes. In experiment 1, sperm motility, abnormalities, membrane integrity, viability, apoptosis status, mitochondrial activity were studied following freeze-thawing. The best quality of frozen-thawed rooster sperm was chosen to be used for the assessment of the hatchability and fertility rate in experiment 2. The significantly higher percentages of post-thawing sperm total and progressive sperm motilities, membrane integrity, viability were observed in 1% soybean lecithin and 20% egg yolk plasma in comparison with 0.5 and 1% soybean lecithin, 10% egg yolk plasma and control, except for 15% egg yolk plasma (P < 0.05). Using 20% egg yolk plasma in the extender improved mitochondrial activity. Supplementation of 1% soybean lecithin and 20% egg yolk plasma into the extender resulted in the least percentages of dead sperm (P < 0.05). Sperm abnormalities and early apoptosis did not differ in various extender supplementations. In experiment 2, higher percentages of hatchability and fertility rate were observed in semen containing 1% soybean lecithin and 20% egg yolk plasma compared with the WEY group. The results showed that supplementation of the rooster sperm extender with 1% soybean lecithin and 20% egg yolk plasma resulted in higher quality of frozen-thawed sperm.

Effects of adding different levels of Glutamine to modified Beltsville extender on the survival of frozen rooster semen.

The aim of the present study was to investigate the effects of l-glutamine on the quality of frozen-thawed rooster semen. Semen samples were collected from eight mature roosters (Ross 308). After initial semen assessments, samples of adequate quality were mixed together and diluted with modified Beltsville extender without l-glutamine (control) and supplemented with 2.5, 5, and 7.5mM l-glutamine. Semen straws were subjected to cryopreservation and evaluated twice at 15-day intervals. After thawing, sperm viability, total and progressive sperm motilities were measured by Eosin-Nigrosine and Computer-Aided Sperm Analysis (CASA), respectively. The results showed that sperm functions decreased on day 30 compared to day 15. The extender supplemented with 5mM glutamine improved (p<0.05) sperm viability, total and progressive sperm motilities compared to other treatments and the control group. The best level of glutamine appeared to be 2.5mM, as it provided the highest sperm membrane integrity and the lowest level of abnormalities. The results of this study suggest that the addition of glutamine to the diluent improves semen quality and using glutamine allows rooster sperm to be frozen for longer.

Effect of vitamin E and selenium nanoparticles on post-thaw variables and oxidative status of rooster semen.

This study was conducted to determine the combined effects of adding vitamin E (VitE) and selenium nanoparticle (Nano-Se) as antioxidant supplements to rooster semen extender for freezing. Semen samples were collected from 12 White Leghorn roosters and pooled. Subsequently, the samples were divided into nine equal groups using modified Beltsville extender. Extenders were supplemented with either two amounts of VitE (5 and 10µg/mL) or two amounts of Nano-Se (1% and 2%) or a combination of both VitE and Nano-Se, and no antioxidants extender (control group). Using 5µg/mL VitE and 1% of Nano-Se improved (P<0.05) total sperm motility (79.28±3.86%), progressive sperm motility (18.03±1.02%), sperm viability (81.46±2.16%) and integrity of the sperm membrane (77.21±2.12%) after the freeze-thawing process compared with control group (54.08±3.86%, 10.96±1.02%, 60.53±2.16%, and 54.47±2.12%; respectively). Also, extenders supplemented with 5µg/mL Vit E or 5µg/mL VitE and 1% Nano-Se had a lesser malondialdehyde (MDA) concentration compared to control extender (1.15±0.32, and 1.29±0.32, respectively). Total abnormal morphology of sperm was decreased (P<0.05) by adding 5 or 10µg/mL VitE alone or in combined with 1% or 2% Nano-Se. Moreover, extenders containing Nano-Se demonstrated greater glutathione peroxidase (GPx) activity compared to other extenders. Catalase (CAT) activity was greater in extender supplemented with 10µg/mL VitE and 2% Nano-Se. Moreover, higher TAC was observed in extenders supplemented with VitE and Nano-Se. It can be concluded that addition of 5µg/mL vitamin E in combined with 1% Nano-Se improved the post-thawing quality and oxidative variables of rooster semen.