The Study Of Hydroxytyrosol Effects On Quality Of Semen During Liquid Preservation At 17? And Artificial Insemination Of Pigs

Artificial insemination technology is the production technology used in more than 99% of the world's pig farms.The quality of breeding and the effect of the effect are the quality of pig semen.The semen storage of most pig farms is carried out at room temperature(17?),and one of the most important problems faced by pig semen during storage at room temperature is the accumulation of reactive oxygen species(ROS).The cumulative excess of ROS is known to attack the sperm membranous system,causing its lipids to peroxidize and damage the sperm plasma membrane.Secondly,ROS as a highly unstable electron-carrying body will attack sperm proteins and DNA.The problem is very likely to cause a drop in semen quality,resulting in problems such as low breeding efficiency,low litter size,stunting,and low birth weight.Therefore,reducing the cumulative damage of ROS during the preservation of pig semen and providing excellent semen quality for breeding is very important for pig production.In this study,we added different concentrations of hydroxytyrosol to the dilution formula during the storage of porcine semen at room temperature,and compared with the conventional dilutions to study the cumulative amount of ROS and the degree of sperm membranous damage during storage.The acrosomal integrity rate,the activity of three antioxidant enzymes in the antioxidant enzyme system,and other indicators to evaluate the effect of hydroxytyrosol on the changes of reactive oxygen species and sperm motility in pig semen,and then determine the hydroxytyrosol in the diluent formulation.The optimum amount to add.After determining the optimal addition amount,the difference in expression of sperm protein with the control group was further analyzed by proteomic analysis at the time of storage of the addition of 120 ?mol/L hydroxytyrosol(optimum concentration)to the third day.Subsequently,the insemination ability of the sperm was evaluated by comparing the different performances of in vitro capacitation and progesterone-induced acrosome reaction in pigs with 120 ?mol/L hydroxytyrosol and ordinary preservation.Finally,the insemination of pig semen in the above two preservation methods was carried out to evaluate the effect of the addition of hydroxytyrosol on the artificial insemination results.Research indicates:1.The ROS content of semen in pig semen stored at room temperature increased exponentially with the prolongation of storage time.The degree of peroxidation of sperm plasma membrane increased,sperm motility decreased,and semen quality decreased.The addition of a low concentration of hydroxytyrosol significantly inhibited the above situation,and when the concentration of hydroxytyrosol was too high,the preservation effect decreased again.According to the test results,the optimum addition amount of hydroxytyrosol in the study was 120 ?mol./L.2.The preservation method of adding low concentration hydroxytyrosol in the dilution formula significantly improved porcine sperm glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),catalase(CAT)The viability(P<0.05),when the addition of hydroxytyrosol was 120 ?mol/L,the enzyme activity of the sperm antioxidant enzyme system was the slowest during the storage period and the total antioxidant capacity was the best.3.According to the results of sperm proteomics analysis,the protein expression of the sperm supplemented with 120 ?mol/L hydroxytyrosol in the dilution formula was significantly different from that of the control(P<0.05).When the differential expression multiple was 1.3 times,the hydroxytyrosol(HT)group expressed 163 up-regulated proteins and 45 down-regulated proteins compared to the control(NC)group.Proteins with differential expression are highly involved in specific cellular processes such as sperm motility and exercise-related processes,sperm redox homeostasis.Through KEGG enrichment analysis,20 metabolic pathways were found to be affected,3 of which were related to sperm energy metabolism and 2 were related to sperm redox homeostasis.The test group has a higher level of expression on sperm energy metabolism,oxidative stress response and other related proteins.4.Compared with the control,there was no significant difference in sperm capacitation rate between the 120 ?mol/L hydroxytyrosol-preserved group(P>0.05),and the acrosome reaction rate was significantly higher than that of the control group(P<0.05).5.Through artificial insemination results,it can be found that the preservation method of 120 ?mol/L hydroxytyrosol in the formula can significantly improve the pregnancy rate of the sow(P<0.01)and the birth weight of the piglets(P<0.05).In summary,the addition of 120 ?mol/L hydroxytyrosol to the diluent formulation can reduce the accumulation of semen active oxygen during the storage of pig semen at room temperature,thereby improving the sperm storage viability and semen quality,and improving the sow after verification by artificial insemination production.Pregnancy rate and birth weight of piglets.