Effect of Diospyros kaki enriched 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<0.000 1) 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<0.003 3) 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 of Diospyros kaki improved bull semen quality post-chilling and post-freezing

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 °C 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 (0.00 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 °C 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<0.000 1) than the control. SEE decreased significantly (P<0.000 1) 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

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<0.000 1) at 6-day post-chilling. In frozen semen, 3%, 4%, 5% and 6% concentrations gave the best significance (P<0.000 1) on sperm motility in comparison with the control. Concentration 1% revealed the highest significance (P<0.000 1) on alive% as compared to the control. Hypo- osmotic swelling test was maintained as the control. Concentration 3% gave the lowest significance (P<0.000 1) 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

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 °C) 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 °C for 5 min to allow uptake of BHT by spermatozoa. The tubes were cooled slowly (approximately for 2 h) up to 5 °C 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

Effects of pomegranate juice in Tris-based extender on cattle semen quality after chilling and cryopreservation

Objective: To study the effect of adding different concentrations of the pomegranate juice (PJ) to the cattle bull semen extender on post-thawing semen quality. Methods: Semen was collected from five cattle-bulls at weekly intervals for 5 weeks at the Semen Freezing Center, General Organization for Vet. Services, Ministry of Agriculture. Semen samples were diluted in Tris-citric acid-egg yolk-fructose extender and divided into six aliquots, the 1st served as control while PJ was supplemented at 10%, 20%, 30%, 40% and 50% in the aliquot 2, 3, 4, 5 and 6 respectively. Diluted semen samples were subjected to cooling and cryopreservation and stored in liquid nitrogen (LN2). Sperm motility in chilled semen (over 10 d) and post-thawing sperm parameters, including individual motility, alive sperm, membrane integrity, and total sperm abnormality were assessed. Results: Obtained results clearly demonstrated that the addition of 10% PJ in the chilled extended cattle semen proved to be beneficial for maintaining sperm motility percentage. On the other hand, the addition of 40% and 50% PJ failed to preserve motility all over the 10 d. Also, supplementation of extender with 10–20% PJ significantly increases the post-thaw motility and viability as compared with control group. Conclusions: Supplementation of bull semen extender with 10% and 20% PJ provides good chilling and improved frozen-thawed semen quality

Effects of different concentrations of sucrose or trehalose on the post-thawing quality of cattle bull semen

Objective: To examine the effect of different concentrations of trehalose or sucrose (50 or 100 or 200 mM) on post-thawed quality of bull semen, cryo-preserved in Tris-citric acid-egg yolk-fructose (TCYF). Methods: Semen samples were diluted in TCYF extender, TCYF + trehalose (50, 100 and 150 mM/L) or TCYF + sucrose (50, 100 and 150 mM/L) to ensure 60 million motile spermatozoa mL-1, cooled slowly up to 5 °C and equilibrated for 4 h. Semen was packed into 0.25 mL polyvinyl French straws. The straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen (LN2) for 10 minutes then dipped in liquid LN2. Frozen straws were thawed at 37 °C for 1 min. The parameters studied were sperm motility, sperm viability, sperm abnormality, sperm membrane integrity (HOST), percent of normal intact acrosome and DNA fragmentation. Results: The output data demonstrated that addition of 50–100 mM of trehalose or sucrose/L TCYF after chilling at 5 °C had significantly (P<0.0001) ameliorated motility, membrane integrity, viability, abnormal morphology, and acrosome integrity % compared to control diluted semen while 50 mM of trehalose/L, and 50–100 mM of sucrose/L to TCYF diluent had significantly (P<0.0001) improved after thawing motility (43.00,% 45.00% and 41.00%, respectively), membrane integrity (67.40%, 67.80% and 69.40%, respectively), life sperm % (70.20%, 69.40% and 71.40% respectively), and acrosome integrity percentages (56.40%, 58.80% and 55.80% respectively) compared to the control tris-base diluent, while diminishing the abnormal sperm morphology (6.20, 3.80 and 3.80 respectively) and DNA fragmentation (3.60%, 3.80% and 3.80% respectively). Besides, the addition of 100 mM of trehalose/L to tris-base diluent has also a promising effect when added to the tris-base diluent concerning the above parameters. Conclusion: It is finally concluded that the addition of 50–100 mM trehalose or sucrose/L to TCYF have a beneficial effect in chilling diluted bull semen, while the use of 50 mM trehalose or 50–100 mM sucrose had their benefits on freezing-thawing of extended semen

Cryopreservation of cattle semen using coconut water extender with different glycerol concentrations

Objective: To investigate the effect of coconut water with a lone concentration and different concentrations of glycerol on chilled and cryopreserved cattle semen characteristics.Methods: Semen was collected from five mature cattle bulls, at weekly intervals for 5 weeks. The ejaculates were pooled and evaluated for dilution processing. Tris citrate egg yolk fructose was used as control treatment for semen, while 50% (V/V) coconut water, 25% (V/V) bi-distilled water and 25% (V/V, 5% anhydrous monosodium citrate) to 20 mL egg yolk and three different concentrations of glycerol (4%, 6% and 8%) were used as coconut water (CW)- glycerol-yolk extenders (CWCG-4, CWCG-6 and CWCG-8). Extended semen was cooled and cryopreserved. Sperm motility%, sperm membrane integrity%, normal acrosome%, live sperm% and total sperm abnormalities% were recorded after equilibrium and after freeze-thawing.Results: The addition of 4% glycerol to coconut water enriched media (CWCG-4) revealed the most effective addition of glycerol on all parameters after equilibrium and after freeze-thawing.Conclusions: Coconut water enriched media with 4% glycerol addition is safe to be used as an extender in bull semen preservation because it is a sterile liquid. So, it can be used without addition of antibiotics to the extender, as antibiotics have to some extent hazardous effect on spermatozoa

Effects of Phoenix dactylifera pollen grains extract supplementation on post-thaw quality of Arabian stallion semen

This study explored the effect of extender supplementation with different concentrations of date palm pollen grain (DPPG) on post-thawed sperm motility, viability index, membrane and acrosome integri-ties in Arabian stallions. Five ejaculates from each of four Arabian stallions were subjected to cryo-preservation with a modified INRA-82, without any supplementation (control) or supplemented with 50, 100, 150, 200 and 250 mg DPPG. After thawing, all samples were maintained at 37 oC, while analyses were performed at 0, 1, and 2 and 3 hours. Sperm motility percentage, viability index, mem-brane integrity percentage and acrosome integrity of each sample were determined by conventional laboratory methods. The addition of 100 mg DPPG resulted in improved maintenance of sperm moti¬lity after 0 and 60 min post-thawing, as compared to the control and other treatment groups. Non-significant effects on viability index were observed after enrichment of extender with 100 and 150 mg DPPG. The addition of 100 and 150 mg DPPG resulted in significant (P<0.0010) improvement in post-thawing membrane integrity (41.33±0.83%; 41.33±2.33%) compared to the controls (34.33±1.55%). These concentrations exerted also a beneficial effect in preserving sperm acrosome integrity (38.33±1.01%; 38.67±1.64%) as compared with the control one (33.33±1.12%). Supple-mentation of modified INRA-82 with 200 mg DPPG failed to maintain sperm motiliy while 250 mg PG has a negative impact on all studied post-thawing semen parameters. In conclusion, adding 100 and 150 mg date palm pollen grain extract to modified INRA-82 seemed useful in the chilling and freezing process of Arabian stallion sperm

Effect of water extract of dates palm (Phoenix dactylifera) on semen characteristics and oxidative status in serum of male New Zealand rabbits under heat stress

Objective: To estimate the effects of the water extract of dates palm (Phoenix dactylifera) (DWE) on sperm quality parameters, testosterone level and serum antioxidants activities of New Zealand rabbits under heat stress.Methods: A total of 30 bucks of New Zealand White rabbits were randomly divided into three equal groups as follows: Group 1 was treated as control group and fed on balanced commercial ration. Groups 2 and 3 were treated with 10 and 20 mL of dates extract substituting water in the early morning before watering and fed on balanced commercial ration. This schedule was performed daily for 5 days/week, for an experimental period of 5 weeks. Fertility parameters such as reaction time, potential of hydrogen ion (pH), mass motility, individual progressive motility %, percentage of live sperm and abnormal sperm (%) were measured. Blood serum testosterone level, serum glutathione reduced, nitric oxide, ascorbic acid and malondialdehyde were also determined.Results: The daily oral administration of 10 mL DWE significantly increased the pH, the mass motility and individual progressive motility % compared to the control group. Although, the consumption of 20 mL DWE significantly (P<0.000 1) increased the live sperm% and decreased the abnormal sperm % compared to the other two treatments. The administration of date extracts (10 and 20 mL) had significantly (P<0.000 1) decreased nitric oxide and glutathione reduced levels compared to the control. On the other hand, it increased significantly the lipid peroxidation, ascorbic acid and testosterone level compared to the control.Conclusions: The aqueous extract of date palm (10-20 mL) could enhance the rabbit bucks fertility and its health performance