Enhancement impact of Moringa oleifera leaves extract–base extender on cryopreservation and fertilization of Barki ram sperms: comparative study with vitamin E and selenium combination

Abstract

This study evaluated the impacts of inclusion of Moringa Oleifera leaves extract (MOLE) in semen extender on rams cryopreserved semen quality and fertilization potential. Forty ejaculates were collected from eight fertile Barki rams, pooled and divided into five groups. The semen extender of the control group was without additives. The semen extender of the second and third groups was supplemented with MOLE at doses of 300 and 600 mg/mL, respectively. The semen extender of the fourth and fifth groups was supplemented with a combination of vitamin E and selenium at doses of 2.5 and 5 mg/mL, respectively. One hundred ten multiparous Barki ewes were artificially inseminated with the semen supplemented without or with MOLE or vitamin E and selenium combination. Inclusion of MOLE in semen extender significantly elevated the motility, viability index, membrane integrity and fertilization capacity of the post-thawed spermatozoa, as well as the activities of semen catalase, total antioxidant capacity, superoxide dismutase, glutathione peroxidase, and glutathione reductase. However, it significantly decreased acrosomal defects and DNA fragmentation of spermatozoa, the activities of semen alkaline and acid phosphatase and the concentration of malondialdehyde compared with the other groups. Similarly, vitamin E and selenium significantly improved the above-mentioned parameters compared to those of the control group. This study indicated that inclusion of MOLE to semen extender improved the quality and fertility of the post-thawed rams' semen through enhancing the activities of the antioxidant enzyme system and decreasing the spermatozoa DNA fragmentation and lipid peroxidation. HIGHLIGHTS Moringa Oleifera leaves extract (MOLE) protected spermatozoa against cryopreservation induced oxidative stress. MOLE enhanced cryopreserved semen quality. MOLE enhanced post-thawed spermatozoa fertilization capacity.

The impact of varying doses of moringa leaf methanolic extract supplementation in the cryopreservation media on sperm quality, oxidants, and antioxidant capacity of frozen-thawed ram sperm

“…In contrast, to control non-treated rams, oral supplementation of rams with MOEE for 64 days increased seminal plasma AA, GPx, and ALP after the treatment but increased GPx, dropped ALP, and did not change AA in the post-thaw semen (Shokry et al 2020). The significant increase of ascorbic acid in the post-thaw semen extenders supplemented with 0.48mg/ml with a slight insignificant increase when 0.56 and 0.64 mg/ml MLME was added in the current study was also noticed when 300 and 600 µg/ ml of moringa leaf extract were added to Barki ram semen (Shokry et al 2021). In contrast to our results where MLME did not increase the concentrations of ALP in the post-thaw semen extenders, it declined it when 300 and 600 µg/ml of moringa leaf extract were added to Barki ram semen (Shokry et al 2021).…”

“…The significant increase of ascorbic acid in the post-thaw semen extenders supplemented with 0.48mg/ml with a slight insignificant increase when 0.56 and 0.64 mg/ml MLME was added in the current study was also noticed when 300 and 600 µg/ ml of moringa leaf extract were added to Barki ram semen (Shokry et al 2021). In contrast to our results where MLME did not increase the concentrations of ALP in the post-thaw semen extenders, it declined it when 300 and 600 µg/ml of moringa leaf extract were added to Barki ram semen (Shokry et al 2021). The significant increase in GPx concentrations when 0.64mg MLME was included is in understanding with including 300 and 600 µg/ml of moringa leaf extricate to Barki rams' semen extenders (Shokry et al 2021).…”

“…The testicular SOD increased and MDA when cryptorchid and non-cryptorchid rats were treated with ethanolic extract of MOLE (Afolabi et al 2013). The significant decrease in the MDA and the increased SOD levels observed herein were noticed also when 300 and 600 µg/MOLE were added to Barki rams' semen (Shokry et al 2021). In contrast, to control non-treated rams, oral supplementation of rams with MOEE for 64 days raised SOD and decreased MDA in fresh seminal plasma and post-thaw semen (Shokry et al 2020).…”

“…The gradual improvement in post-thaw sperm motility and viability observed in the current study with increasing MLME concentrations from 0.48, 0.56, to 0.64 was reversed when 0.5, 5.0, and 10.0 moringa seed methanolic extracts were added to ram semen (Carrera-Chávez et al 2020) indicating that the improvement in post-thaw semen is concentration-dependent as >0.56 and >0.65 mg are more preferable and higher concentrations that were associated with lowered post-thaw antioxidant capacity and sperm membrane damage (Carrera-Chávez et al 2020). However, in Barki rams, the increasing moringa leaf extract from 60 to 600 µg in the semen extenders could increase post-thaw sperm motility and viability but from 900 and 1200 µg/ml could reduce them to values lower than control, indicating that this improvement is dose dependent (Shokry et al 2021). In contrary, the oral supplementation of Barki rams MOLE for 64 days did not alter sperm motility, viability, abnormal sperm, membrane functional integrity, and DNA fragmentation (Shokry et al 2020).…”

“…In another study, MOLE supplemented orally to Barki rams did not improve spermatozoa motility and viability over 64 days of the administration, but it improved their antioxidant status (Shokry et al 2020). Higher post-thaw sperm motility and viability were reported following fortification of Barki ram freezing extender with MOLE (Shokry et al 2021). In rams, Moringa oleifera seed methanolic extract improved post-thaw semen motility and antioxidant capacity (Carrera-Chávez et al 2020).…”