Effect Of Sperm Storage Tubules Secretions From Pre-Layer Hen On Cockerel Sperm Clumping And Motility
Material type: Book ; Format:
Publisher: 2004 Dissertation note: Artificial insemination (AT) in the poultry industry has considerable importance because of better results in fertility and hatchability. Increasing male utilization in artificial insemipation depends upon the optimum use of semen by suitable diluting media to increase the volume of ejaculate and to preserve fertility.
In the present study, the effect of sperm storage tubules secretions on percentage motility and extent of clumping of sperms was noticed. An optimum osmotic pressure 375 mOsm with pH 7.0 was used to preserve the cockerel's semen at 5°C. A total of 20 meat Breeder cockerels were randomly selected. After providing 10-days sexual rest, they were trained for semen collection by abdominal massage technique. Three birds failed to produce good quality semen. These birds were removed from the study.
Semen from seventeen Meat Breeder Cockerels was pooled. After
macroscopic evaluation, the pooled semen was divided into 4 groups.Group A was diluted with Modified Van Wambeke diluent with addition of sperm storage tubules secretions. Group B was diluted with the above diluent without SST secretions. Group C was diluted with saline solution with addition of SST secretions while group D was diluted with saline solution without SST secretions. These four groups were stabilized at 375 mOsm osmotic pressure in pH 7.0 and stored at 5°C. The diluted semen samples were examined for percentage motility and extent of clumping. After 72 hours of semen storage (5°C), group A showed significantly (P<0.05) higher motility as compared to groups B, C and D.
The extent of clumping was higher (P <0.05) in group D as compared to groups A, B and C. However, group A showed less (P <0.05) clumping upto 64 hours as compared to groups B, C and D.
The results of the present study suggested that at 375 mOsm, pH 7.0 the cockerels semen stored at 5°C diluted wiih Milk based extender and saline solution with addition to SST secretions proved to be suitable for short-term preservation of Meat Breeder Cockerel semen.
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Effect Of Cold Shock On Frozen-Thawed Spermatozoa Of Buffalo & Cow Bulls
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; Literary form:
Publisher: 2005 Dissertation note: Effect of cold-shock on frozen-thawed bovine spermatozoa was measured in terms of motility, viability, plasma membrane and acrosome . integrity. Single ejaculates each from seven Nili-Ravi buffalo bulls and 7 Sahiwal cow bulls were processed for freezing. Three straws of 0.5ml (for each parameter) from each bull were thawed and pooled at 3rC before and after cold-shock at 4°C for 2 min. Individual motility was observed by phase contrast microscope before and after cold-shock. Before and after cold-shock, sperm viability, plasma membrane and acrosome integrity were evaluated by supravital stain, hypo osmotic swelling test and normal acrosomal reaction, respectively. Two hundred sperm were counted for each parameter before and after cold-shock. Comparison of before and after cold shock values showed that cold shock had no effect (P>0.05) on motility (64.0±1.88 vs 60.4±2.56), viability (141.1±4.39 vs 124.5±8.03), plasma membrane integrity (83.51±5.82 vs 71.19±4.31) and acrosome integrity (148.3±2.36 vs 141.1 ±2.85) of buffalo bull spermatozoa. Comparison of before and after cold shock values of cow bull spermatozoa indicated that cold shock had significant (P<0.05) effect on motility (59.2±1.05 vs 41.9±.12), viability (140: 14±2.94 vs 90.5±2.73), and plasma membrane integrity (62.3±4.28 vs 47.24±3.71) but no effect on acrosome integrity (147.9±2.21 vs 140.6±2.40). In conclusion, cold shock had significant detrimental effect on cattle bull spermatozoa but no effect on buffalo bull spermatozoa.
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Effect Of Osmotic Pressure On The Membrane Integrity Of Frozen-Thawed Buffalo Bull Spermatozoa
Material type: Book ; Format:
Publisher: 2005 Dissertation note: In the first experiment, semen from seven Nili-Ravi buffalo bulls was used to study plasma membrane integrity of freshly collected (raw) and frozen-thawed sperm using the hypo-osmotic swelling test (HOST). For this purpose, percentage motility, integrity of plasma membrane and acrosome was assessed by a phase contrast microscope, HOST plus eosin-nigrosin staining and normal apical ridge test. respectively. 50fJI each of raw and frozen-thawed semen was mixed with 500fJI of 50, 100, 150, 190 or 250 mOsm hypo-osmotic treatments of sodium citrate plus fructose and incubated at 37°C for 1 h. Integrity of sperm plasma membrane was assessed before and after hypo-osmotic treatments to estimate the extent of damage for each hypo-osmotic treatment. In raw semen, the number of swollen sperm was higher (P<0.05) at 50, 100, 150 and 190 mOsm as compared to 250 mOsm. A positive but non-significant correlation (P>0.05) was found between percentage of swollen and live sperm at 100, 150, 190 mOsm. Similarly, a positive but non-significant correlation (P>0.05) was found between percentage of swollen sperm and motility at 50, 100, 150, 190 and 250 mOsm. In frozen-thawed semen, the number of swollen sperm w.as higher (P<0.05) at 50 and 100 mOsm as compared to 150, 190 and 250 mOsm and this number decreased significantly (P<0.05) and gradually from 82.6±5.99 at 150 mOsm to 69.7±5.49 at 190 mOsm and 42.6±4.07 at 250 mOsm. A positive but non-significant correlation (P>0.05) was found between percentage of swollen and live sperm and between percentage of swollen sperm and motility at 50, 100 150, 190 and 250 mOsm. The number of sperm with intact acrosome did not differ (P>0.05) in raw and frozen-thawed semen among treatments. Percentag~ motility in raw semen was highe~' (81%) as compared to frozen-thawed semen (60%). Damage to plasma membrane'was higher (P<0.05) at 50 mOsm (59% raw vs. 70% frozen-thawed) as, compared to other hypo-osmotic treatments, while minimum damage occurred at 250 mOsm (4.1% raw vs. 9.7% frozen-thawed). In the second experiment, hypoosmotic swelling test (HOST) plus eosin-,nigrosin staining and normal apical ridge test (NAR) were used to determine integrity of plasma membrane and acrosome of raw, diluted (cooled to 5°C) and frozen-thawed sperm. Semen from seven bulls was used in this study. For diluted and frozen-thawed sperm, three straws were pooled at 37°C. Percentage motility of raw, diluted and frozen-thawed sperm was assessed using a phase contrast microscope. 501-11 each of raw, diluted and frozen-thawed sperm was mixed with 5001-11 of 50, 100, 150, 150, 190 or 250 mOsm hypo-osmotic treatments of sodium citrate plus fructose and incubated at 37°C for 1 h. Total number of intact (live) sperm of raw, diluted and frozen-thawed semen was assessed before HOST. Diluted sperm showed higher (P<0.05) swellings of plasma membrane at 50
and 100 mOsm than raw sperm. Similarly, swellings of diluted sperm were' higher (P<0.05) than frozen-thawed sperm. Swellings of raw sperm were significantly higher (P<0.05) at 100, 150, 190 and 250 mOsm than frozenthawed sperm. A significant decrease (P<0.05) was found among percentage motility of raw (81±1.57), diluted (69.6±2.24) and frozen thawed (60.1 ±1.34) sperm. Live sperm were higher (P<0.05) in raw (174.4±7.33) and diluted semen (175.6±3.76) as compared to frozen-thawed semen (142.3±4.84). Although integrity of the acrosome of raw, diluted and frozen-thawed sperm did not differ (P>0.05), significan~ variation was found within bulls. In conclusion, fresh and frozen-thawed sperm behaved differently to HOST and the number of swollen sperm was higher in raw as compared to frozenthawed semen. Plasma membrane' integrity of raw and diluted sperm was compromised during freezing and thawing. However, freezing had no effect on acrosome integrity. Moreover, 100, 150 and 190. mOsm were found suitable to perform HOST.
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