Effect ofDiospyros kakienriched extender on cattle bull sperm parameters and conception rate

Objective:To explore the effect of Diospyros kaki on cattle spermatozoa during chilling and cryopreservation.Methods: Five milliliter of blended Persimmon (Diospyros kaki) flesh was added to 45 mL TCF to obtain 10% stock solution. Kaki enriched extender (KEE) was prepared by adding to TCF in concentrations 0.0/5.0 mL (control, 0%), 0.5/4.5 mL (1%), 1/4 mL (2%), 1.5/3.5 mL (3%), 2.0/3.0 mL (4%), 2.5/2.5 mL (5%), 3.0/2.0 mL (6%), 3.5/1.5 mL (7%), 4.0/1.0 mL (8%), 4.5/0.5 mL (9%) and 5.0/0.0 mL (10%) to obtain a final volume 5 mL in each tube. Whole egg yolk was added to each tube to obtain KEE with 20% egg yolk (KEEY), all tubes were centrifuged to get rid of debris. Semen was added to the supernatants in other tubes. Extended semen was subjected to evaluation (motility, alive sperm and intact sperm membrane (HOST) %) in both chilled and cryopreserved semen. Conception rate was carried out.Results:Sperm motility was significantly (P<0.0001) kept high after 11 d of chilling with the concentration 1%, 2%, 3%, 4%, 5% as compared to the control (41.67±1.67, 41.67±1.67, 40.00±0.00, 41.67±1.67 and 41.67±1.67, respectively) and also non-significantly kept high at the other concentrations up to 9 d of chilling. Addition of KEE had significantly (P<0.0033) improved post thawing sperm motility % with the concentrations 1, 2, 3, 4, 5 and 6% as compared to the control (51.67±5.27, 55.00±3.16, 48.33±1.05, 45.00±3.96, 57.00±2.50, 55.00±5.00 and 43.33±5.11 respectively).While the other concentrations exhibit no effect. Addition of KEE maintained alive sperm%, abnormalities% and % of intact spermatozoa membranes (HOST %) as good as the control with all concentrations of kaki used in our study. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 2%, 4% and 6 % KEE in cattle.Conclusion: It could be concluded that some concentrations ofDiospyros kaki improved bull semen quality post-chilling and post-freezing.

Effect of tris-extender supplemented with various concentrations of strawberry (Fragaria spp.) on bull semen preservability

Objective:To evaluate effect of tris-extender supplemented with various concentrations of strawberry (Fragaria spp.) on bull semen preservability.Methods: Pooled bull semen were extended with tris-citrate-fructose egg yolk diluent (control, 0% strawberry) and various concentrations of tris strawberry (TSB) (1%-6%) to achieve 60 million motile spermatozoa per milliliter. Extended semen were subjected to semen freezing protocol. Semen assessment including motility, alive%, abnormality%, intact sperm membrane (hypo-osmotic swelling test) and conception rate were carried out for both chilled and frozen semen. Results: Results showed that sperm motility after chilling was enhanced in groups treated with various concentrations of TSB from 1% to 5% and exhibited higher significance (P<0.0001) at 6-day post-chilling. In frozen semen, 3%, 4%, 5% and 6% concentrations gave the best significance (P<0.0001) on sperm motility in comparison with the control. Concentration 1% revealed the highest significance (P<0.0001) on alive% as compared to the control. Hypo-osmotic swelling test was maintained as the control. Concentration 3% gave the lowest significance (P<0.0001) considering abnormality%. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 5% and 6% in cattle.Conclusions: It is concluded that 1%-5% concentrations of TSB ameliorate bull semen characteristics after chilling, and 3%-6% concentrations of TSB improve bull semen characteristics after freezing. Higher conception rate exists in 5% and 6% concentration of TSB.

Impact of silymarin enriched semen extender on bull sperm preservability

Objective:To explore the effect of silymarin on bull spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk diluents, purified silymarin powder (obtained from the milk thistle silybum marianum), purchased from Unipharma, Al Obour city, Egypt, was soaked in Tris-citric acid-fructose diluent for 48 h at 10℃ making a stock solution (70 mg/mL), from this stock solution we obtained concentrations of 0.18 mg/mL, 0.36 mg/mL, 0.54 mg/mL, 0.72 mg/mL, 0.90 mg/mL in addition to the control (0.0 mg/mL) reaching a final volume of 5 mL in each tube. Egg yolk was added to each tube to obtain silymarin enriched semen extender (SEE) with 20% egg yolk, cooled slowly up to 5℃ and equilibrated for 4 h. Semen was packed into 0.25 mL polyvinyl French straws (IMV, France). After equilibration periods, the straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen for 10 min and were then dipped in liquid nitrogen. Extended semen was subjected to evaluation (motility, alive %, abnormality %, intact sperm membrane (HOST)% and conception rate) in both chilled and frozen semen.Results: Table 1 revealed that Sperm motility of the concentrations 2, 3 and 4 after 8 d of chilling were significantly (P<0.02) higher than control. Sperm motility of the concentration 2 (45.00%±2.89%) after 9 d of chilling was higher than control and the other concentrations. Addition of SEE in concentration 1 and 2 gave post thawing sperm motility as high as the control (47.50±2.81 and 45.00±2.58, respectively) while other concentration have lower effects on motility as compared to the control. Addition of silymarin improved post thawing alive% and was significantly higher (P<0.0001) than the control. SEE decreased significantly (P<0.0001) the % of post thawing abnormal sperm in concentration 3 and 4 (11.83±0.65 and 16.00±0.58, respectively). SEE improved significantly (P<0.018) the % of post thawing intact spermatozoa membranes (HOST%) in concentrations 2, 4 and 5 (71.17±0.83, 71.83±0.91 and 75.00±3.42 respectively) (Table 2).Conclusion:It could be concluded that silymarin as a natural additive to semen extenders improved preservability in both chilled and frozen bull semen.

Preservability of bull spermatozoa in Tris-egg yolk extender enriched with different concentrations of butylated hydroxytoluene

Objective:To explore the effect of BHT on cattle spermatozoa during cooling and cryopreservation.Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk (TCFY) diluent considered as control (0 BHT) and different concentrations of BHT (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mM were prepared in ethanol in prewarmed (37℃) test tubes. The ethanol was allowed to evaporate so that, a thin crystallized layer of BHT was deposited on the inner surface of the tubes. Then extended semen was added into the tubes and incubated at 37℃ for 5 min to allow uptake of BHT by spermatozoa. The tubes were cooled slowly (approximately for 2 h) up to 5℃ and equilibrated for 4 h. After equilibration, semen freezing process was carried out. Extended semen was subjected to evaluation (motility, alive sperm, intact sperm membrane (HOST) % and acrosome integrity) in both cooled and cryopreserved semen. Results:The result revealed that sperm motility of post-cooled spermatozoa improved (P<0.05) by the use of BHT concentrations (1, 2 and 3 mM) in Tris semen extender if compared to the control (85.00±1.09), (83.33± 0.63), (81.67± 0.63) and (78.33± 0.63), respectively. Alive sperm percent was significantly higher in all concentrations of BHT. Sperm abnormalities percent were significantly lower in concentrations of BHT 1 and 2 (11.2±0.2), (11.8±0.2)and (13.4±0.4), respectively. Sperm membrane integrity were significantly higher in BHT concentrations (1, 2, 3, 4 and 5 mM). It is exhibited that improved sperm motility in post-thawed frozen semen in the concentrations of BHT (1, 2, 3 and 4 mM) if compared to the control. The sperm membrane integrity were significantly improved at all concentrations of BHT. Acrosome integrity was significantly higher at BHT concentration 1 mM (81.80±0.57) and (76.00±2.05), respectively.Conclusions: It could be concluded that some concentrations of BHT improved bull semen quality post-cooling and post-freezing.