abouzar najafi

Artificial insemination is one of the important management tools to control reproductive activities and increase the productivity of the livestock industry. This technique can play an important role in genetic improvement even in small herds. Sperm preparation is of particular importance for reproductive techniques, such as artificial insemination, but many harmful factors, such as temperature shock and oxidative stress, reduce sperm quality and directly affect sperm fertility during sperm preparation and storage. Therefore, the addition of protective substances and antioxidants in the diluent is considered essential in maintaining sperm quality during the cooling and freezing stages. Curcumin is a yellow substance obtained from the root of turmeric, and its biochemical structure contains a phenolic ring and a beta-di-ketone part in its structure. It can neutralize free radicals, but it has low solubility in hydrophilic environments. To solve this problem, the nanoliposome method has been proposed. Using this method can solve the problem of curcumin's low solubility in aqueous solutions and, additionally, curcumin nanoparticles can be guided to exert their effects in a completely targeted manner inside the target cell. On the other hand, sucrose protects sperm from cold shock by first rinsing sperm cells and maintaining the osmotic pressure of the diluting medium. This study aimed to investigate the effects of separate and mixed use of different levels of curcumin nanoparticles and sucrose sugar in ram epididymal sperm diluent during storage at 5 ˚C at different cooling times.

The present study was conducted in the Animal Physiology Laboratory of the Department of Animal and Poultry Sciences, Aburihan Agricultural Campus, the University of Tehran. The testicular tissues were prepared from a slaughterhouse on the day of the experiment and transported to the laboratory with special flux in less than an hour. Sperm samples were collected from the epididymal tail of the ram testicular tissue in the laboratory. Sperm was sampled after initial evaluation, with progressive motility of more than 75% and abnormality of less than 10%, and were added to the diluent at 37 °C. In this study, the diluent consisted of Tris (27.1 g/L), citric acid (14 g/L), fructose (10 g/L), egg yolk (20%), and glycerol (7%), with a pH of around 6.2-7.8. The osmolarity was set at about 310-320 milliosmoles. Sucrose sugar was obtained from the Sigma Aldridge Company, and curcumin nanoparticles were prepared from pure curcumin powder (Sigma Aldridge) by the nanoliposome method and using rotary devices, a homogenizer, and a prop sonicator. A DLS device was used to examine the particle size of curcumin nanoparticles (less than 80 nanograms). The experimental groups included 25 and 50 μM of nanoparticles of curcumin (NC), sucrose (S) at 100 and 150 mM separately, combined levels (25NC100S, 25NC150S, 50NC100S, and 50NC150S), and a group without these additives (control), which were added to the diluent at 37 °C. The diluents containing different levels of the treatments were refrigerated in 15 ml falcones and inside a water container at the same temperature, which gradually reached a temperature of 5 ˚C in about 2 hours. After stabilization at this temperature, total motility parameters, progressive motility using the KASA system, the viability with the eosin-nigrosin method (16.7 g of eosin, 100 g of nigrosin, and 29 g of citrate buffer in 1 L of double-distilled water), membrane integrity by the HOST solution method (9 g of fructose and 4.9 g of sodium citrate in 1 L of double-distilled water with an osmolarity of 100 milliosmoles), and the percentage of sperm abnormalities using Hancock's solution (5 62/L of 37% formalin, 150 ml of saline solution, 150 ml of a buffer solution, and 500 ml of double distilled water) were evaluated at 6, 12, 24, and 48 hours.The experimental results were analyzed in a completely random design by SAS software (9.2) and the GLM procedure at a significance level of P < 0.05. The means of the treatments were compared with Tukey's test.

The use of 25NC100S in the diluent improved the parameters of total motility, progressive motility, viability, and integrity of ram epididymal sperm membrane in all evaluated times compared to the other groups, especially the control group (p < 0.05). No significant differences were observed between the treatments in the percentage of sperm abnormalities at 6 and 12 hours (p > 0.05). At 24 and 48 hours, however, the 25NC100S concentration significantly reduced the percentages of sperm abnormalities compared to the other concentrations (p < 0.05). Compared to the control group, adding different levels of curcumin nanoparticles and sucrose sugar separately significantly improved the evaluated parameters, but the best performance was observed in the combined treatment, indicating the synergistic role of these two compounds.

The results of the present study demonstrate that using a mixture of curcumin nanoparticles with sucrose in the diluent can protect sperm against oxidative damage and cold shock. The antioxidant effects of curcumin nanoparticles and the protective effects of sucrose sugar can result in positive effects on the quality of ram epididymal sperm during cooling. Therefore, it is recommended to use the 25NC100S level in ram sperm diluents.

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.

Japanese quail has a rapid growth rate (3 to 4 generations per year) and is relatively resistant to many diseases. Due to its high productivity, it has gained importance as an animal model in biological and genetic studies worldwide. Nutrition is a key factor that affects production efficiency in quails. Japanese quails require a diet that contains high-quality protein and a balanced profile of amino acids. Branched-chain amino acids (leucine, isoleucine, and valine) not only contribute to protein synthesis but also play a role in other metabolic activities. Among the branched-chain amino acids, valine is a limiting amino acid in corn-soy-based diets. Protein is the most expensive component of the diet and has a significant impact on production costs in poultry. Nowadays, reducing the protein level in the diet has garnered attention for cost reduction in farming and for mitigating environmental pollution resulting from ammonia emissions and nitrogen excretion in the poultry industry. However, reducing the protein level in the feed without proper supplementation of amino acids can lead to decreased feed intake, reduced production levels, and changes in social behavior in birds. Published reports indicate that adding crystalline amino acids to low-protein diets in meat birds can have similar performance to high-protein diets at different growth stages. In most cases, valine becomes limiting with reduced protein levels in the diet. Considering the important metabolic roles of branched-chain amino acids, especially valine, in rapidly growing birds, the aim of this experiment was to investigate the effects of using diets containing different digestible valine levels on the performance, growth traits, and histology of the liver and magnum in Japanese quails during the growth period.

All experimental procedures for the care and use of animals in the present study were approved by the animal care committee of the University of Tehran. Thousand one-day-old quails were given 5 dietary treatments, such that each treatment was replicated 5 times and 40 birds per replicate in a completely randomized design, during 1–42 days of age. Experimental diets were formulated to meet nutrients recommendation of growing quails with different levels of dietary digestible valine concentration (0.75, 0.85, 0.95, 1.05 and 1.15 %) in diets with a low protein (17.7-17% protein). The experimental diets provided 2,9 kcal metabolizable energy per gram. Other nutrients were provided based on the recommended nutritional requirements for growing quails. Other nutrients were formulated based on the recommended nutritional requirements for growing quails. The experimental diets were adjusted for two age periods: 1-21 days and 22-42 days. The birds received 24 hours of light until three days of age, followed by 23 hours of light and one hour of darkness daily. The parameters investigated in this experiment included growth performance, feed consumption, conversion ratio and carcass performance. The bird weights were measured at the beginning of the experiment and on days 21 and 42. Feed consumption on days 21 and 42 was calculated based on the difference between the amount of feed provided and the remaining feed. The weight of dead birds was used to correct the amount of feed consumption. On day 42, one male quail and one female quail were randomly selected from each replicate and killed after 6 hours of starvation. Additionally, after sampling the liver and magnum tissues, their histomorphological characteristics (nuclear diameter of hepatocytes, diameter of sinusoids in the liver tissue; epithelial height, inner muscle thickness, outer muscle thickness, gland diameter, and gland depth in the magnum tissue) were investigated.

During the breeding period, the quails that received the diet containing 0.95% and 1.05% digestible valine had higher body weight gain (BWG) and feed conversion ratio (FCR), respectively. Feed intake (FI) and mortality were not affected by the treatments. Dietary treatments did not have any significant effect on the carcass characteristics (both male and female). However, the carcass yield was higher in male quails and the relative weight of breast and intestines was higher in female quails. In the treatments containing 1.05% and 1.15% of valine, the diameter of Liver cell nucleus and the diameter of liver hepatocytes were the highest, which indicates liver damage. The volume of sinusoids in female birds increased with increasing valine levels in the diet; So that in birds that received diets containing levels higher than 0.95% valine was more than birds fed with rations containing 0.75% valine. Based on the results of this study, different levels of valine in diets containing 17% crude protein had a significant effect on the histology of the magnum tissue (epithelial height, inner muscle thickness, outer muscle thickness, gland diameter), although only the depth of the glands showed a tendency for significance with 0.85% valine.

Based on the results, increasing the level of digestible valine in low protein diets up to 0.95% improves growth performance and feed conversion ratio in growing quails. However, it seems that the use of high levels of digestible valine in low protein diets (more than 0.85%) may lead to tissue damage in the liver and have a negative effect on growth performance. Further studies are recommended in this field.

In the present experiment, the effects of different concentrations of rosemary extract(RE) spray on fertile eggs on the first day of incubation on embryonic parameters and quality of hatched chicks were investigated. Seven treatments were tested in a completely randomized design with five replicates, including five treatments of different concentrations of rosmarinic acid(RA) (0.5,0.1,1.5,0.2, and 2.55 mg/ml of RA) and two treatments of distilled water (with and without spray) on the first day of incubation. The results showed that the weight of the yolk sac of hatched chicks from eggs sprayed with a concentration of 2.55 mg/ml of RA was significantly lower than those from eggs treated with distilled water(P<0.05).There was no significant difference in hatchability percentage and embryonic mortality between unsprayed and sprayed eggs with distilled water compared to RE. The use of RE spray on egg caused a reduction in the body surface temperature(BST) of hatched chicks compared to those from unsprayed eggs(P<0.05). The length of the intestine of hatched chicks from eggs sprayed with 1.5 mg/ml of RA was greater than that of unsprayed and distilled water-sprayed eggs(P<0.05). Finally, the results showed that the spray of RE, with a concentration of 2.55 mg of RA, resulted in a heightened absorption of yolk sacs in hatched chicks compared to those eggs sprayed with distilled water. Moreover, an longer intestinal length was observed using a spray of 1.5 mg of RA, accompanied by a decrease in body surface temperature of hatched chicks in comparison to eggs without any spray.

The number of teat is an important trait in relation to the maternal ability of high litter size sheeps, it is important to know the genetic function and controlling positions of this trait in the genomes of different species. High litter size domestic animals are potentially dependent on colostrum-derived immunoglobulins due to placental structure in the early hours of life. Therefore, the number of teat plays a critical role when there are more offspring born at birth than the number of teats. In some cases, particularly the number of the extra teat and even their position is very important. A method to identify new loci and confirm existing QTL is through genome-wide association studies (GWAS). QTL assisted selection and genomic regions affecting the production traits have been considered to increase the efficiency of selection and improve production performance. Genome wide association studies typically focus on genetic markers with the strongest evidence of association. However, single markers often explain only a small component of the genetic variance and hence offer a limited understanding of the trait under study. A solution to tackle the aforementioned problems, and deepen the understanding of the genetic background of complex traits, is to move up the analysis from the SNP to the gene and gene-set levels. In a gene-set analysis, a group of related genes that harbor significant SNP previously identified in GWAS, is tested for over-representation in a specific pathway. The present study aimed to conduct a genome wide association studies (GWAS) based on Gene-set enrichment analysis for identifying the loci associated with teat number trait using the high-density SNPs.

Hu sheep, a descendant of Mongolian sheep, is a famous lambing breed in China. In ewes of the Hu sheep, TT (individuals with two normal teats) and MT (individuals with two normal teats and one or two supernumerary teats) account for 76–62% and 38–24% respectively. For MT ewes, the supernumerary teats are smaller than the normal teats, but some can produce milk. In the current study, we analyzed the complex genetic mechanism of differences in teat number using single-locus GWAS in a total of 160 Hu sheep. The gene set analysis basically consists of three different steps: the assignment of SNPs to genes, the assignment of genes to functional categories, and finally the association analysis between each functional category and the phenotype of interest. Genome wide association study was performed with birth weight and biometric traits using GEMMA software. Using the biomaRt2 R package, the SNP were assigned to genes if they were within the genomic sequence of the gene or within a flanking region of 25 kb up- and downstream of the gene. For the assignment of the genes to functional categories, the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway databases were used. The GO database designates biological descriptors to genes based on attributes of their encoded products and it is further partitioned into 3 components: biological process, molecular function, and cellular component. The KEGG pathway database contains metabolic and regulatory pathways, representing the actual knowledge on molecular interactions and reaction networks. Finally, a Fisher’s exact test was performed to test for overrepresentation of the significant genes for each gene-set. The gene enrichment analysis was performed with the goseq R package. In the next step, a bioinformatics analysis was implemented to identify the biological pathways performed in BioMart, Panther, DAVID and GeneCards databases.

Gene set enrichment analysis has proven to be a great complement of genome-wide association analysis (Gambra et al., 2013; Abdalla et al., 2016). Among available gene set databases, GO is probably the most popular, whereas KEGG is a relatively new tool that is gaining ground in livestock genomics (Morota et al., 2015, 2016). We had hypothesized that the use of gene set information could improve prediction. However, neither of the gene set SNP classes outperformed the standard whole-genome approach. Gene sets have been primarily developed using data from model organisms, such as mice and flies, so it is possible that some of the genes included in these terms are irrelevant for meat production. A better understanding of the biology underlying meat production specifically, and an advance in the annotation of the ovine genome, can provide new opportunities for predicting production using gene set information.In this research, 7 SNP markers on chromosomes 3, 5, 7, 8, 12, 14 and 17 located in CDH11, NUMB, FGF2, ESR1, LGR5, INSR and PTGS2 genes were identified. Some of the found genes, are consistent with some of the previous studies related to teat number traits. According to pathway analysis, 11 pathways from gene ontology and biological pathways were associated with teat number (P˂0.05). Among these pathways, Blastocyst development, Mesenchymal cell development, Developmental growth involved in morphogenesis, Muscle cell differentiation and AMPK signaling pathway have important functions in development of mammary gland and activation of the AMPK signaling pathway. Finally, it is worth noting that our gene-set enrichment analysis was conducted using a panel of SNP obtained from a single marker regression GWAS, which relies on a simplified theory of the genomic background of traits, without considering the joint effect of SNP. Hence, other approaches (e.g., GWAS exploring SNP by SNP interactions) might provide a better basis for biological pathway analysis.

In total, this study supported previous results from GWAS of teat number, also revealed additional regions in the sheep genome associated with these economically important trait. Using these findings could potentially be useful for genetic selection in the breeding programs.

The present study aimed to conduct a genome wide association studies (GWAS) based on Gene-set enrichment analysis for identifying the loci associated with growth traits using the 50K arrays. For this purpose, phenotypes records and genotypic data related to birth weight (BW), weaning weight (WW), six month weight (6MW) and 12 month weight (12MW) were obtained from 240 Hu sheep. Genome wide association study was performed with growth traits using TASSEL software. Using the biomaRt2 package R the SNP were 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. By Gene set enrichment analysis, the biological pathways and candidate genes of regulation of skeletal muscle cell proliferation  and actin filament polymerization (MYOD1 and IGFBP4), regulation of anatomical structure morphogenesis, regulation of muscle system process and cell junction (BMP2, THBS1 and MYH10), regulation of skeletal muscle tissue development and regulation of anatomical structure size (FGF2, ANGPTL4, TGFBR3 and ACTR3), anatomical structure morphogenesis, positive regulation of ossification and cellular response to growth factor stimulus (MMP7, ACTA2, EGR1 and MYBPC3), for BW, WW, 6MW and 12MW were identified, respectively. The detected candidate genes played an important role in muscle structure development, glucose metabolism, Osteoclast differentiation, body size, skeletal muscle cell differentiation and regulation of ion calcium. These novel candidate genes identified in this study may be useful targets for clarifying our understanding of the molecular basis of growth traits in sheep. These results can provide new insight for future research into the genetic architecture of growth traits in sheep breeding program.

Type traits describing the skeletal characteristics of an animal are moderately to strongly genetically correlate with other economically important traits in cattle including fertility, longevity and carcass traits. The present study aimed to conduct a genome wide association studies (GWAS) based on gene-set enrichment analysis for identifying the loci associated with conformation traits using the 50K arrays.

Genome wide association study was performed with biometric traits using GEMMA software v. 0.98. Using the biomaRt2 R package R the SNP were assigned to genes if they were within the genomic sequence of the gene or within a flanking region of 25 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, 12 SNP markers on chromosomes 1, 3, 5, 7, 8, 10, 13, 16, 17, 22, 23 and 25 located in PLCB1, PTBP1, TMEM130, PLCB4 (BL), TGFBR3, SYN3, TRAK1 (WH), GLP1R, HMGA1 (CG) and CAPN3, MYOG and MYO18B (CD) genes were identified. Some of the genes that were found are consistent with some previous studies related biometric traits. According to pathway analysis, 17 pathways from gene ontology and biological pathways were associated with biometric traits (p˂0.05). Among these pathways, muscle structure development, anatomical structure formation involved in morphogenesis, anatomical structure homeostasis, Osteoclast differentiation, skeletal muscle cell differentiation and calcium signaling pathway have important functions in development of skeletal muscle, glucose homeostasis, osteogenesis process, regulation of ion calcium and activation.

The finding of this research confirms the results of previous studies from genomic scanning related to type traits, also revealed additional genomic regions, could be useful for genetic selection in the cattle breeding programs.

Red blood cells play an essential role in the oxygen transport and the immune system. Moreover, hematologic parameters are an important clinical indicator of various diseases including anemia and metabolic syndrome.

The present study aimed to conduct genome-wide association studies (GWAS) based on gene-set enrichment analysis to identify the loci associated with hematological traits using 630K arrays.

For this purpose, the phenotype records included 498 genotyped Alpine Merino sheep were used for red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and RBC volume distribution width coefficient of variation (RWD_CV). Genome-wide association study was performed with hematological traits using TASSEL software. Using biomaRt2 R package R, SNP was assigned to genes. GO, KEGG, DAVID, and PANTHER databases were used to assign the genes to functional categories.

11 SNP markers on chromosomes 1, 2, 3, 6, 8, 10, 11, 14, and 20 located in TRPC4, SPAT1, TMCC2 (RBC), KRT26, GPLD1, EPAS1 (HGB), RAC2, HSPD1, PDGFRA (HCT) and BBS1, HAG1, PIK3R3, STXBP5, FCER1G  (MCH, MHCH, RWD_CV) genes were identified. Based on the pathway analysis, 17 pathways from gene ontology and biological pathways were associated with hematological traits (P˂0.05). The pathways have important functions in the development and differential of red blood cells, hypoxia, adaptation process, environmental stress, and platelet activation.

In total, this study supported previous results from the GWAS of hematological traits, and also revealed additional regions in the sheep genome associated with important traits, using these findings could be potentially useful for genetic selection in the breeding programs.

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.

Oxidative stress is an important factor in poor sperm function that causes morphological changes and oxidative damage. Various studies have shown that antioxidants can reduce oxidative damage during freezing. In fact, antioxidants reduce, eliminate, and inactivate free radicals. Many compounds, such as some plant extracts, some industrial chemicals, and some amino acids, can act as active antioxidants and neutralize free radicals. luteolin is an important member of flavonoids which is present in glycosylated form in plants and studies have shown that luteolin has antioxidant properties. The aim of this study was to reduce lipid peroxidation and minimize oxidative stress by adding different levels of luteolin in the freezing medium.

Semen samples were collected from 15 roosters at 28 weeks of age. Sperm collection from roosters was performed by back-abdominal massage method. To eliminate individual effects after initial evaluations, the samples were mixed together and used for cryopreservation. After extending the samples and adding the levels of 0 (control), 0.25, 0.5, 0.75, 0.1 and 1.25 μM luteolin to the base extender (lake extender), sperm were added at 1:20 ratio. The test samples were cooled to 4 ° C and then frozen in liquid nitrogen. The samples were thawed at 37 ° C and then the motility parameters were measured by CASA computer system, survival by eosin nigrosine test, plasma membrane integrity by host test, morphology by Hancock test and the amount of produced malondialdehyde. Then the data were analyzed by completely randomized design.

The results showed that the addition of 0.75, 1 and 1.25 μM levels significantly increased the total motility and the levels of 0.75 and 1 increased the progressive mobility and VAP. In addition, 1 μM of luteolin caused a significant increase in VSL parameter (p <0.05). The results showed that levels of 0.75 and 1 μM significantly increased membrane integrity, and also level of 0.75 μM of luteolin decreased malondialdehyde content and sperm percentage with abnormal morphology (p <0.05).

The results of this study showed that the addition of luteolin in the extender during freezing and thawing can have a positive effect on improving the quality of rooster sperm and subsequently increase the total and progressive motility after freezing and thawing and also the level of 0.75 μM could be the best performance among different levels added to the extender.

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.

Understanding the genetic control of temperament as a complex trait and correlated with economic traits is one of the breeding goals in beefcattle industry. The aim of the current study was genome wide association studies (GWAS) based on Gene set enrichment analysis fordetecting the loci associated with temperament traits in Brahman cattle breed. Therefore, 1370 Brahman cattle and phenotype recordsassociated with temperament traits including Exit velocity, Pen Score, and Temperament Score were used. The evaluation of genome-wideassociation was carried out using PLINK package 1.90. The gene enrichment analysis was performed by the goseq R package for identifyingbiological pathways of nearby genes in selected candidate regions and finally, GO, Metacyc, KEEG, Reactome and panther databases wereapplied for bioinformatics analysis. By Gene set enrichment analysis, the biological pathways and candidate genes of neurotransmittersecretion (NRXN3 and CACNG3), Dopamine Neurotransmitter Release Cycle (PPFIA2), regulation of neuron projection development(GRID2), neuron projection (SLC8A1 and KCNQ2), Axonal growth inhibition (RTN4R), Neurotrophin signaling pathway (MAP2K2,MAP3K5 and PSEN1) and Focal adhesion (TLN2) were identified. The detected candidate genes played an important role in differentiationof synapse, neurotransmitters, neurological diseases and disorders, oxidative and environmental stresses, hormone receptors and glucosehomeostasis. Considering the confirmation of the previous region of genome wide association and the identification of new genomic regions,the findings of this study can be useful in the genetic selection of higher production cattle through calm animals.

The purpose of this study was to investgate resveratrol nano-protected antioxidant on rooster sperm. In this study, resveratrol was used in the forms of nano-protected (liposome and NLC) and plain in the semen extender. Immediately after the semen collection and for primary evaluations, for eliminating of individual effects the semen collected from 15 roosterswere pooled and added to the extender. Extending was done at 37 °C at a dilution ratio of 1:20. Different concentarions of resveratrol (20, 40 or 60 μM) were added to the extender in three forms of plain, NLC, and liposome. After diluting the semen, gradual cooling was carried out for 3 h at 4 °C. After the cooling process, the kinetic parameters (CASA), membrane integrity (HOST), viability, lipid peroxidation (MDA), abnormal sperm, were evaluated. The results at 24 and 48 hours at a temperature of 4°C indicate that the use of 40 μM resveratrol, resveratrol loaded in the liposome, and resveratrol loaded in the NLC improved the motility, progressive sperm motility, viability, and membrane integrity compared to the control group. The results of this experiment indicated that 40 μM resveratrol, resveratrol loaded in liposome and resveratrol loaded in NLC improved most of the evaluated parameters.

In order to improve the quality of ram semen after freezing-thawing process, different levels of ethanol extract of Bilberry plants were investigated. In this study five rams were used for semen collection twice a week by an artificial vagina. In order to eliminate the effects of the individual, the semen samples were pooled. Different levels of Bilberry extract (0, 100, 150, 200 μl/ml) were added to the Tris- Egg yolk based extender. Following cooling and freezing of semen samples, they were stored in liquid nitrogen until evaluation. After freezing-thawing, motility parameters were evaluated using the Computer-assisted sperm analysis system (CASA), the viability of sperms by Eosin-Nigrosin staining, membrane integrity using the hypo-osmotic swelling test, sperm abnormality by Hancock solution and lipid peroxidation by measuring malondialdehyde concentration. The results showed that the addition of 150 μl of Bilberry extract improved sperm parameters following freezing-thawing process compared to the control group (P