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This study was conducted to assess the influence of supplementing Tris-extender with Coenzyme Q10 (COQ10) as an antioxidant on sperm characteristics of Egyptian buffalo and cattle. This study was carried out at Animal Production Research Station, El-Gemmezah, belonging to Animal Production Research Institute, Egypt. Five sexually mature buffalo (4 y and 400-450 kg) and Friesian (3 y and 350-400 kg) bulls were used in this study. Semen was collected twice weekly for five successive weeks. Only ejaculates with ≥70% mass motility were pooled and diluted at 37°C with Tris-extender containing 0, 20 and 30 μM of CoQ10 in T1, T2 and T3, respectively. Equilibration period was at 5°C for 4 h and semen was packed in 0.25 ml French straws frozen in liquid nitrogen-196 o C. Stored straws were thawed individually at 37°C for 30 s. Semen was evaluated for sperm characteristics including percentages of progressive motility, livability, abnormality, curled tail and acrosomal damage of spermatozoa. Enzymatic activity of AST, ALT and LDH was determined in seminal plasma of post-thawed semen. Results showed that there were no species differences in all sperm characteristics studied, except for increasing (P<0.05) percentage of abnormality and damage acrosome spermatozoa in post-thawed semen of cattle compared with buffalo bulls. All sperm characteristics were improved (P<0.05) by 30 μM COQ10 supplementation T3 in post-diluted, post-equilibrated and post thawed semen as compared to control without supplementation T1 in buffalo and cattle semen. Activity of all enzymes reduced (P<0.05) in seminal plasma of both species. In conclusion, CoQ10, as antioxidant, at a level of 30 μM in Tris-extender is able to enhance most sperm characteristics including mobility, livability, plasma membrane integrity, with decreasing sperm abnormalities and it has strong protective power against acrosomal damage of buffalo and cattle spermatozoa.

This study was designed to evaluate the possibility of using the optimal level of Moringa oleifera leaf aqueous extract (10, 15 or 20%) with 90, 85 or 80% saline (MOLE) as a semen extender of bull semen in comparing with three extenders, including Tris-egg yolk (TEY) without or with vitamin C at a level of 0.9 mg/ml (TEY+VC), and camel skim milk (CSM). Semen was collected from five Friesian bulls (350-450 kg LBW and 3-4 years old), twice a week for 5 weeks using artificial vagina. After ejaculation, mass motility was evaluated and only those of ≥70% were used in this study. Ejaculates were pooled, divided into 4 replicates and prepared for freezing. Semen was extended in TEY (control), TEY+VC, (CSM) and 15% MOE+85% saline (based on the best results of MOLE level). Semen was diluted, equilibrated at 5 o C (4 h) and placed in liquid nitrogen (LN) at-196 o C. Frozen semen stored for one month at least was warmed at 37 o C for 30 sec. Semen evaluation for percentages of individual motility (IM), live sperm (LS), sperm abnormality (SA), acrosomal damage (AD) and hypo-osmotic swelling test (sperm cells with curled tail, CT) was estimated in diluted, equilibrated and thawed semen, while percentage of head to head agglutination (HHA) was determined in post-thawed semen only. Activity of AST, ALT, LDH and total antioxidant activity were determined in seminal plasma of post-thawed semen. Conception rate (CR) was recorded for Friesian cows inseminated with semen frozen with the four types of extenders. Results showed that percentages of IM, LS, SA, AD and CT in post-diluted and post-equilibrated semen, beside all previous characteristics and percentage of HHA in post-thawed semen showed the best results (P<0.05) in CSM, moderate in 15% MOE and TEY+VC, and the poorest in TEY. Enzyme activities of AST, ALT and LDH in post-thawed seminal plasma were the lowest (P<0.05) in CSM, followed by 15% MOE, while total antioxidants activity was higher (P<0.05) for TEY+VC, CSM and 15% MOE than for TEY, being the highest for 15% MOE. Artificial insemination of estrous synchronized cows with semen extended by different types of extenders indicated that conception rate of cows was significantly (P<0.05) the highest for Friesian cows inseminated with post-thawed semen extended with CSM (87.5%), moderate with both TEY+VC and 15% MOE (75%) and the lowest with TEY(62.5%). Extender containing 15% MOE showed the cheapest cost (4.80 L.E./100 ml) in comparing with CSM (6.45 L.E./100 ml), TEY+VC (13.95 L.E./100 ml), and even the conventional TEY extender (13.85 L.E./100 ml). In conclusion, extender containing 15% Moringa oleifera leaf extract and 85% saline solution could be successfully used, as a source of antibiotics and antioxidants, in extenders of frozen bull semen, being with the cheapest cost as compared to tris-egg yolk or camel skim milk extenders.

This study was conducted at Animal Production Research Station, El-Gemmezah, belonging to Animal Production Research Institute, Egypt, in cooperation with Department of Animal Production, Faculty of Agriculture, Mansoura university. Aim of the current study was to evaluate the possible effects of leptin and melatonin as a protective additive in Tris-extender on frozen semen quality of buffalo bulls. Five sexually mature Egyptian buffalo bulls (aged 4 to 5 years) with an average live body weight of (450-500 kg). All bulls were fed on the same diet. Semen was collected, twice a week for 4 wk (40 ejaculates for each bulls), using artificial vagina. The collected ejaculates (mass motility of more than 70%) were pooled, evaluation, divided into 7 replicates and frozen. Semen was extended in Tris-egg yolk extender as unsupplemented (T1), or supplemented with leptin (10, 20 and 50 ng/ml in T2, T3 and T4, or melatonin (10-3 M, 10-6 M and 10-9 M in T5, T6 and T7), respectively. Semen was diluted , equilibrated at 5 0 C for 4 h and frozen in liquid nitrogen. After storage for 4 weeks, frozen straws were thawed at 37 0 C for 30 sec. Semen was evaluated in post-dilution, post-equilibrated and post-thawed semen. Various sperm parameters including percentage of individual motility (IM), dead sperm (DS) and sperm abnormality (SA) were determined. In the seminal plasma of thawed semen, activity of AST, ALT and LDH was measured. Total of 12 buffalo cows in heat were inseminated by semen extended with T1, T3 and T5 to determine conception rate (CR). Pregnancy rate was measured by rectal palpation on day 50 after (AI). The obtained results show that IM and SA percentages improved (P<0.05) in T3 and T5. Percentage of DS improved (P<0.05) in all treatments as compared to control. T3 showed the best (P<0.05) results concerning sperm characteristics studied in postdiluted as compared to other additives and T1. IM, DS and SA percentages showed the best (P<0.05) results in post-equilibrated semen (T3), followed T5 in comparison with other additives and control. IM, DS and SA improved (P<0.05) in post-thawed semen diluted (T3) or T5, being better with T3 more than with T5. Enzyme activity of AST, ALT and LDH reduced (P<0.05) in seminal plasma of post-thawed semen diluted with all additives as compared to control, being the lowest (P<0.05) in T3. Buffalo cows inseminated with semen (T3) showed higher (P<0.05) CR (91.6%) than T5 (75.0%) or control (T1, 66.6%). In conclusion, supplementation of 20 ng/ml leptin and 10-3 M melatonin to the freezing semen extender improved semen quality and reduce cryodamage of the buffalo bull spermatozoa.

Low fertility in buffaloes with frozen-thawed semen is attributed to sperm damage that is related to formation of ice crystals during freeze/thaw process. This study was conducted to evaluate the freezability of buffalo bull semen extended with four types of extenders, including citrate egg-yolk (CEY, control), CEY with ethanolic Moringa Olifera Leaf Extract (eMOLE) as replacement of antibiotics (CEYm), eMOLE and eMOLE with conventional antibiotic (eMOLEa) with or without Coenzyme Q10 as antioxidant. Semen was subjected to calculate progressive motility, livability, abnormality, acrosome status and integrity of sperm membrane in semen after extending, equilibrating and thawing processes. Four sexually mature buffalo bulls (4-5 y and 400-450 kg LBW) were used for semen collection, and semen was collected twice weekly for 4 successive weeks. Semen ejaculates of ≥70% as mass motility percentage were pooled and extended at 37°C by the four types of extenders without or with 30 µM Co-Q10). Results cleared that progressive motility, livability, abnormality, acrosomal damage and plasma membrane integrity percentages improved (P<0.05) in buffalo semen after dilution, equilibration and thawing, while activity of AST, ALT and LDH in seminal plasma of post-thawed semen reduced (P<0.05) by extension of semen with eMOLE as a new extender. All sperm parameters and enzyme activity were maintained by replacing antibiotics in CEY extender with 4 ml eMOLE. All sperm parameters were improved (P<0.05) and enzyme activity was maintained by adding 30 µM of coQ10 as compared to free extenders. Fertility rate of buffalo cows inseminated with semen cryopreserved in extender (eMOLE+coQ10) was the highest (91.6%), followed by eMOLE without coQ10 (83.3%) and the lowest for control CEY (66.6%). In conclusion, eMOLE as a new extender, is promising for the extension and preservation of buffalo semen. Also, adding 4 ml of eMOLE could be a good substitute for the antibiotic component of CEY as a conventional semen extender. Moreover, supplementing semen extenders with antioxidant addition (30 µM CoQ10) is important to improve function of spermatozoa and sperm fertility of cryopreserved semen of buffaloes.