International Journal of Veterinary Sciences Research

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No. 3

Effects of Degasified Extender on Quality Parameters of Cryopreserved Bull Spermatozoa

Pages: 70-78
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Effects of Degasified Extender on Quality Parameters of Cryopreserved Bull Spermatozoa

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DOI: 10.18488/journal.110/2015.1.3/110.3.70.78

Citation: 4

Megha Pande , N. Srivastava , J.S. Rajoriya , S.K. Ghosh , J.K. Prasad , S.S. Ramteke

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Megha Pande , N. Srivastava , J.S. Rajoriya , S.K. Ghosh , J.K. Prasad , S.S. Ramteke (2015). Effects of Degasified Extender on Quality Parameters of Cryopreserved Bull Spermatozoa. International Journal of Veterinary Sciences Research, 1(3): 70-78. DOI: 10.18488/journal.110/2015.1.3/110.3.70.78
One of the most important factors contributing to poor quality semen post thaw has been reported to be oxidative stress. Continued interaction of spermatozoa with dissolved oxygen in extender renders detrimental effect by generating reactive oxygen species during processing. This formed the basis to investigate the effects of removal of oxygen from the Tris-Glycerol-Egg Yolk (TG-EY) extender on the post-thaw semen quality. Semen ejaculates (n=26) collected from three cross bred bulls with mass activity of ≥3 and progressive motility of ≥70% were split in two fractions (control and treatment) to obtain 80 x 106 progressively motile sperm/mL of extended semen. The TG-EY extender composed of 3.028 g of Tris, 1.675 g of citric acid, 1.25 g of fructose, 7.0 mL of glycerol, 10 mL of egg yolk, 105 IU Penicillin G Sodium and 105 µg Streptomycin for 100 mL of deionized water for control and degasified (freeze-pump-thaw cycling method using liquid nitrogen) for treatment. Semen was processed and filled in 0.25 mL polyvinyl chloride straws, sealed and cryopreserved for at least 2 week before evaluation. Assessment of post-thaw motility, viability, acrosome and plasma membrane integrity of spermatozoa were evaluated after thawing at 37°C for 30s. Study revealed a significant (viability and acrosome integrity, p<0.05) and highly significant (post-thaw motility and plasma membrane integrity, p<0.01) improvement in all the semen quality parameters in degasified as compared to control group. This study showed that degasification of extender prior to their use can serve to improve semen quality to highly acceptable levels.
Contribution/ Originality
This study shows importance of degasification of extender in cryopreservation protocol. The assays used to evaluate effect of degasification were currently in vogue. Fewer studies were available to evaluate the effect of degasification in cattle sperm cryopreservation. The paper contributes by showing the importance of degasification on sperm cryosurvival.

Cholesterol Content of Bull Spermatozoa Alters Survival at Ultra-Low Temperatures

Pages: 63-69
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Cholesterol Content of Bull Spermatozoa Alters Survival at Ultra-Low Temperatures

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DOI: 10.18488/journal.110/2015.1.3/110.3.63.69

N. Srivastava , S.K. Srivastava , S.K. Ghosh , Amit Kumar , Megha Pande , P. Perumal , Y.K. Soni

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N. Srivastava , S.K. Srivastava , S.K. Ghosh , Amit Kumar , Megha Pande , P. Perumal , Y.K. Soni (2015). Cholesterol Content of Bull Spermatozoa Alters Survival at Ultra-Low Temperatures. International Journal of Veterinary Sciences Research, 1(3): 63-69. DOI: 10.18488/journal.110/2015.1.3/110.3.63.69
Though cryopreservation bestows many advantages, it decreases tolerance to stress of temperature variations, reducing survival of spermatozoa to a great extent. A gradual reduction in cholesterol content of spermatozoa during cryopreservation was reported. Thus the objective of this investigation was to evaluate relationship of cholesterol content of spermatozoa with surviving ability, biochemical integrity of membrane and in vitro fertility (IVF) of spermatozoa following cryopreservation. From each ejaculate (n=12) aliquot was taken for evaluation of semen quality parameters and thereafter it was processed for cryopreservation. Cholesterol content, viability, motility and biochemical integrity of membrane (Hypo osmotic swelling, HOS response) of spermatozoa at fresh, pre-freeze and frozen-thaw stages and IVF parameters at frozen-thaw stage were evaluated. Values were fitted in prediction equation to predict  cryosurvivability and biochemical integrity of spermatozoa membrane. Study indicated that cholesterol content of fresh spermatozoa can be used to predict viability at pre-freeze and frozen-thaw stages of cryopreservation protocol with medium level of accuracy (p<0.05). Cholesterol content of fresh, pre-freeze and frozen-thaw spermatozoa can be used to predict HOS response with medium level of accuracy (p<0.05) at respective and forthcoming stages. However cholesterol content of spermatozoa was not found to be significant predictor of individual motility and in vitro fertility (penetration percent and penetration index). This study revealed evidence that cholesterol content of fresh, pre-freeze as well as frozen-thaw bull spermatozoa can be a good predictor of viability and biochemical integrity of spermatozoa membrane following preservation at ultra low temperature.


Contribution/ Originality
This study contributes by way of showing importance of sperm cholesterol in cryopreservation. The assays used were currently in vogue. The equation is new one and not many studies have been done on this aspect. The paper contributes in emphasizing that sperm cholesterol content has an important bearing on cryosurvival.

Effect of Using Different Pre-Storage Warming Times on Hatchability of White Hisex Breeders’ Eggs

Pages: 54-62
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Effect of Using Different Pre-Storage Warming Times on Hatchability of White Hisex Breeders’ Eggs

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DOI: 10.18488/journal.110/2015.1.3/110.3.54.62

Atif, A. H. , Sayda, A. M. , ElBeeli M. Y. M , Elfadil, A. A. , Fawgia, Sir E. S.

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Atif, A. H. , Sayda, A. M. , ElBeeli M. Y. M , Elfadil, A. A. , Fawgia, Sir E. S. (2015). Effect of Using Different Pre-Storage Warming Times on Hatchability of White Hisex Breeders’ Eggs. International Journal of Veterinary Sciences Research, 1(3): 54-62. DOI: 10.18488/journal.110/2015.1.3/110.3.54.62
It is well known that commercial hatcheries set their eggs after days of storage which increases the incubation duration, decreases hatchability, chick quality and growth performance. The objective of this experiment was to study the effect of different pre-storage warming (PRESW) times on hatchability, embryonic mortality and chick grades of White Hisex layer breeders’ eggs. A total of 1200 eggs were collected from a flock at 67 weeks of age. Eggs were divided into four groups of 300 eggs each according to their warming time (0 hour as control, 2 hours, 4 hours and 6 hours, respectively). These groups were further subdivided into four replicates. of 75 eggs each and were assigned to the completely randomized design (CRD). Eggs were incubated at 37.5°C. All eggs after warming were stored for two days in a cooler at 18°C and a relative humidity of 75%, they were then incubated in (Pas Reform) setter for 18 days and hatcher for three days. At the end of the hatching process hatched chicks were graded (1st and 2nd grade chick); pipped - hatched eggs were then removed and counted. The remaining unhatched eggs were broken to determine fertility and embryonic mortality. Results indicated that pre-storage warming of hatching eggs at 37.5oC for 4 hours significantly (P ≤ 0.01) reduced early dead embryos and total unhatched eggs. The first grade chicks were significantly (P ≤ 0.01) higher in pre-storage warming eggs. It is concluded that 4 hours PRESW improved hatchability percentage as it decreased embryonic mortality percentage, increased the number of saleable first grade chicks which by far increases profits.
Contribution/ Originality
This study is one of very few studies which have investigated the effect of pre-storage of hatching eggs after laying on hatchability, embryonic mortality and chicks’ quality in the region and it is the first one in Sudan.