Effect Of Chlorogenic Acid On Boar Semen Normal Temperature Prervation

As an auxiliary reproductive technology,artificial insemination has been widely used in pig production.The preservation of porcine semen has become the key to improve the quality of semen preservation as the key step of artificial insemination.Protection of spermatozoa from oxidative damage of reactive oxygen species and contamination of bacteria during preservation has become the key to improving semen preservation quality.Chlorogenic acid,a common antioxidant,antibacterial and antiviral component in Chinese herbal medicine,has been widely used in food processing,cosmetics and pharmaceutical industries.In this study,chlorogenic acid was added to the semen diluent for semen preservation at room temperature.Firstly,high throughput sequencing was used to detect the species and level of bacteria during the preservation of Yorkshire semen,and the bacteriostatic and antioxidation ability of chlorogenic acid was verified in vitro;secondly,he conventional indexes,antioxidant indexes and bacteriostatic effects of different concentrations of chlorogenic acid on semen preservation at room temperature were investigated,and the optimum concentration of chlorogenic acid was selected.Finally,the substitution effect of chlorogenic acid on gentamicin was studied,and the substitution effect of chlorogenic acid was verified with gentamicin.The main results obtained in this study are as follows,1.Yorkshire semen were mainly Gram-negative bacteria in the preservation process at room temperature,mainly for Bordetella,Delfactrium,Serratia,etc.;chlorogenic acid concentration is 0.02 ~ At 0.10 g/L,the antibacterial effect against Escherichia coli and Staphylococcus aureus was lower than 0.25 g/L gentamicin(P < 0.05),but the antioxidant performance was stronger than that of vitamin C(P < 0.05).2.The preservation effect of chlorogenic acid 0 to 0.10 g/L in BTS diluent was changed first and then decreased.The addition of 0.06 ~ 0.08 g/L chlorogenic acid significantly increased sperm viability,exercise performance,plasma membrane integrity,and ATP levels(P < 0.05).3.The addition of 0.08 g/L chlorogenic acid significantly increased the total antioxidant capacity and superoxide dismutase(SOD)levels in semen,and significantly decreased the production of malondialdehyde and reactive oxygen species(P < 0.05).4.The bacterial content of pig semen was negatively correlated with the concentration of chlorogenic acid,but the antibacterial effect was lower than 0.25 g/L gentamicin(P < 0.05).0.08 g/L chlorogenic acid can replace the addition of 50% gentamicin,and the combination of the two can increase the storage time of pig semen at room temperature for 7 days.In summary,porcine semen is mainly Gram-negative bacteria during normal temperature preservation;0.08 g/L chlorogenic acid in BTS diluent can significantly increase sperm viability,exercise performance,membrane integrity,antioxidant capacity and antibacterial ability.However,the antibacterial effect was less than 0.25 g/L gentamicin.By adding 0.125 g/L gentamicin and 0.08 g/L chlorogenic acid in combination,semen preservation could be increased to 7 days.