Evaluation of the effects of hesperidin on fresh and frozen-thawed semen quality using two different cryopreservation methods in Simmental bull

Abstract

In the industry of bull semen freezing centers, one-step and two-step semen dilution protocols are two standard and well-known methods in semen freezing process. As the freezing/thawing processes cause detrimental effects on sperm function, the addition of antioxidants can improve sperm characteristics. Hesperidin (Hesp) is an antioxidant used as the male reproductive protective agent. Therefore, the aim of this study was to investigate two different dilution methods, as well as to evaluate Hesp supplementation influence on sperm characteristics in fresh and frozen thawed semen. Semen samples were collected from 12 Simmental bulls. Two separate examinations were conducted in, with and without Hesp supplementation groups. Statistical analysis was performed by an independent t-test, Mann Whitny test, MANOVA and ANOVA tests. In comparison to the one and two-step dilution protocols without Hesp supplementation, the two-step dilution showed greater cryoprotective potential. In the Hesp supplemented group, each semen sample was divided into six equal parts for experimental groups (dilution step method/µM of Hesp). In the both one and two step dilution protocols, significant improvements were detected in semen motility parameters by Hesp administration. Also, oxidative stress status was reduced in seminal plasma of Hesp treatment groups. Interestingly, in comparison with Hesp dosage, 1µM was shown to have greater semen cryoprotective potential. In conclusion,

The influence of L‐proline and fulvic acid on oxidative stress and semen quality of buffalo bull semen following cryopreservation

“…A standard Tris‐based extender (Tris: 2.66 g per 100 mL, citric acid: 1.47 g per 100 mL, glucose: 0.63 g per 100 mL, egg yolk: 20% (v/v), glycerol: 7% v/v, penicillin: 1 mg per 100 mL, streptomycin: 1 mg per 100 mL and pH: 6.8), according to previous research (Topraggaleh et al., 2014), was used as base extender in this study. In each replicate, semen samples ( n = 3) were pooled and equally divided to 12 parts and each part was diluted to a final concentration of 15 × 10 6 spermatozoa/mL (Tahmasbian et al., 2022) with one of the following extenders including: Group 1: Control (Tris‐based extender without antioxidant); Lp‐10: Tris‐based extender + L‐proline (10 mM); Lp‐20: Tris‐based extender + L‐proline (20 mM); Lp‐40: Tris‐based extender + L‐proline (40 mM); Lp‐60: Tris‐based extender + L‐proline (60 mM); Lp‐80: Tris‐based extender + L‐proline (80 mM) (Li et al., 2021); FA‐0.2: Tris‐based extender + fulvic acid (0.2% w/w); FA‐0.5: Tris‐based extender + fulvic acid (0.5% w/w); FA‐0.8: Tris‐based extender + fulvic acid (0.8% w/w); FA‐1.1: Tris‐based extender + fulvic acid (1.1% w/w); FA‐1.4: Tris‐based extender + fulvic acid (1.4% w/w); FA‐1.7: Tris‐based extender + fulvic acid (1.7% w/w) (Xiao et al., 2018). The straws were put in a refrigerator set at 4˚C for 2 h and then semen was frozen according to the following protocol: from + 4°C to −12˚C at a rate of – 4˚C /min, from – 12˚C to – 40˚C at a rate of – 40˚C/min and from – 40˚C up to – 140˚C at a rate of – 50 ˚C/min.…”

“…1.47 g per 100 mL, glucose: 0.63 g per 100 mL, egg yolk: 20% (v/v), glycerol: 7% v/v, penicillin: 1 mg per 100 mL, streptomycin: 1 mg per 100 mL and pH: 6.8), according to previous research (Topraggaleh et al, 2014), was used as base extender in this study. In each replicate, semen samples (n = 3) were pooled and equally divided to 12 parts and each part was diluted to a final concentration of 15 × 10 6 spermatozoa/mL (Tahmasbian et al, 2022) (Xiao et al, 2018). The straws were put in a refrigerator set at 4˚C for 2 h and then semen was frozen according to the follow-…”

Effect of various hesperidin and naringin addition levels on specific sperm indicators and the integrity of their DNA as measured by the comet assay in aged broiler breeder males

“…The majority of seminal fluid properties deteriorated along with the storage of avian spermatozoa at refrigerator temperature for 72 hours, according to several studies. Hesperidin and naringin have been added to semen extenders for bulls and rams, and this practice has been found to be effective [19,20]. In order to better understand how hesperidin and naringin affect the qualities of semen as well as the degree of genetic damage in broiler breeder rooster sperm, this study was created.…”