• Asian-Australasian Journal of Animal Sciences
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• 제22권5호
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• pp.626-635
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• 2009

An experiment was conducted to investigate the effect of Butylated Hydroxy Anisole (BHA), Butylated Hydroxy Toluene (BHT), Pentoxifylline (PTX), Theophylline (TPY) and Theobromine (TBR) on cold protection ability of Murrah buffalo semen at room ($22-25^{\circ}C$) and refrigerated temperature ($4-7^{\circ}C$). Each semen sample was divided into six parts of equal volume and sperm concentration; the first was kept as a control and the remaining five were treated with BHA, BHT, PTX, TPY or TBR. Sperm motility, abnormal spermatozoa, live-dead count, hypo-osmotic swelling and acrosomal integrity were studied at room and refrigerated temperature for various incubation periods viz.; 0, 4, 8, 12 and 24 h at room and 0, 12, 24, 36, 48, 60 and 72 h at refrigerated temperature. Significant improvement in sperm motility, live-dead count, hypo-osmotic swelling and acrosomal integrity were observed in BHT, PTX and TPY fortified extender at room and refrigerated temperature for various incubation periods. From the present study it could be concluded that cold protection ability of buffalo semen can be improved through the addition of BHT followed by PTX and TPY.

• Asian-Australasian Journal of Animal Sciences
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• 제16권10호
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• pp.1424-1428
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• 2003

Present investigation was conducted to determine the post-thaw sperm motility and acrosomal damage of filtered and non-filtered frozen semen of Murrah buffalo bulls. Twenty semen ejaculates (from four Murrah buffalo bulls collected at weekly interval) were diluted in Tris egg yolk glycerol extender and divided into two parts. One was filtered through sephadex G-100 column and the other portion was kept as such (non-filtered). Both fractions were frozen in liquid nitrogen ($-196^{\circ}C$) by the standard method developed in the laboratory. After 24 h of freezing, non-filtered and filtered semen samples were thawed at $37^{\circ}C$ for 1 min. These samples were incubated at $37^{\circ}C$ in a water both. The different seminal characteristics i.e. percent progressive sperm motility, live and abnormal spermatozoa and spermatozoa with damaged acrosome were assessed at hourly interval till they remained motile. The filtered frozen and thawed semen showed significantly (p<0.05) high sperm viability and acrosomal integrity as compared to non-filtered semen.

• 한국가축번식학회지
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• 제3권1호
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• pp.41-47
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• 1979

This experiment was carried out to clarify the methods of the F-body test in human and the B-body test in buil and hog. The effect of pH and albumin concentration on the migration of X- and Y- sperm was also investigated. The results obtained were summarized as follows: 1. In the human semen, the frequency of sperm in which an F-body is visible was different by the fluorochrome. Namely, in case of quinacrine mustard, the F-body frequency was 48.8∼43.4 percent (average 49.6%), and in case of quinacrine dihydrochloride, that was 40.7∼50.8 percent (average 42.0%). 2. The frequency of a, pp.rance of B-body was 43.4${\pm}$1.3 percent in bull semen, and 45.5${\pm}$0.7 percent in hog semen. 3. A, pp.arance of B-body in bovine semen was increased due to duration of time after washing till 12 hours. 4. Separation of X- and Y- spermatozoa using diluents with different hydrogen ion concentration was impossible. 5. A, pp.arance of B-body separated in medium with 6, 10 and 20% ovalbumin was 51.1${\pm}$2.4, 50.6${\pm}$2.5 and 58.2${\pm}$3.0 percent, respectively, and those values were significiantly higher (p<0.01) than corresponding control values.

• Asian-Australasian Journal of Animal Sciences
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• 제16권2호
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• pp.165-171
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• 2003

Sixty semen ejaculates collected at weekly interval from four Murrah Buffalo bulls over a period of seven months (Nov.1999 to May 2000) were used in the present study. Three buffer medium (sodium citrate, TES and Tris) were used for soaking of sephadex. Three grades of sephadex (G-15,G-100, and G-200) were used for preparation of columns. Columns of three different height (one, two and three cm) were used for separation of semen. Twenty semen ejaculates were used in each project. In the first experiment each semen ejaculates was divided into four parts. One part was kept as control and other three parts were passed thought one cm column of sephadex G-15 prepared in three different buffers. There was significant (p<0.05) increase in percent progressive sperm motility and percent live spermatozoa and decrease in percent abnormal spermatozoa and percent spermatozoa with damaged acrosome as well as sperm numbers after filtration through all the three columns. Sperm quality obtained in the filtrate of column prepared in Tris buffer was better in comparison to other two buffers. So the Tris buffer was used in the second trial. Twenty semen ejaculates were used in this experiment. Each semen ejaculate was divided into four parts. One part was kept as control (non-filtered) and other three parts were passed through columns of different grade of sephadex (G-15, G-100 and G-200). Progressive sperm motility and live sperm percentage improved significantly while decline in percent abnormal spermatozoa and percent spermatozoa with damaged acrosome and sperm concentration was observed after filtration through all the columns as compared to control (non-filtered) semen. Since post filtration quality of semen was better in the sephadex G-100 column, therefore it was selected for the next experiment. In third experiment, Tris buffer and sephadex G-100 were used for preparing columns of different height (one, two and three cm) and twenty semen ejaculates were filtered. The quality characteristics of semen (percent progressive sperm motility, percent live spermatozoa and sperm concentration) after filtration through one cm column were significantly (p<0.05) higher than after filtration through columns of two and three cm height. However non -significant (p>0.05) difference due to height of columns was observed for percent abnormal and percent damaged acrosome but 1 cm column comparatively gave better result than of 2 and 3 cm column height.

• Asian-Australasian Journal of Animal Sciences
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• 제27권11호
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• pp.1554-1561
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• 2014

Present study analyzed the changes in peripheral blood testosterone concentrations and testicular cytogram in relation to age and semen quality in crossbred males. Three different age groups of crossbred males viz. bull calves (6 months, n = 5), young bulls (15 months, n = 5) and adult bulls (4 to 6 years, n = 8) were utilized for the study. Testicular fine needle aspiration cytology technique was used to quantify testicular cytology and their indices. Peripheral blood testosterone concentrations were measured using enzyme-linked immunosorbent assay method. Semen samples collected from adult bulls were microscopically evaluated for quality parameters. Mean peripheral blood testosterone concentrations in bull calves, young bulls and adult bulls were $2.28{\pm}0.09ng/mL$, $1.42{\pm}0.22ng/mL$ and $5.66{\pm}1.08ng/mL$ respectively, and that in adult bulls were significantly different (p<0.01) from young bulls and bull calves. There was no significant difference between the proportion of different testicular cells in bull calves and young bulls. Between young and adult bulls, significant differences (p<0.01) were observed in the proportion of spermatocytes, spermatozoa, and sperm: Sertoli cell ratio. The proportions of Sertoli cells showed a significant difference (p<0.01) between the three age groups. The number of primary spermatocytes had a positive correlation with peripheral blood testosterone concentrations in bull calves (r = 0.719, p<0.01). Number of Sertoli cells per 100 germ cells was negatively correlated with blood testosterone concentration in young bulls (r = -0.713, p<0.01). Among different semen parameters in adult bulls, ejaculate volume (r = 0.790, p<0.05) had positive relationship, and sperm motility had significant negative correlation (r = -0.711, p<0.05) with testosterone concentrations. The number of Sertoli cells and Sertoli cell index had a positive correlation with various semen quality parameters (p<0.001). Results of the present study conclude that number of Sertoli cells and Sertoli cell index are good indicators of semen quality, but peripheral blood testosterone concentrations may not have a direct relationship with various seminal attributes in crossbred bulls.

• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1188-1195
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• 2001

A tetraparental chimeric bull was successfully produced by aggregating bovine IVF embryos of F1 (female Holstein${\times}$male Japanese Black) and F1(female Japanese Brown${\times}$male Limousin) and culturing in vitro without the zona pellucida at Yamaguchi Research Station in Japan. In the microsatellite genotyping, 12% (28/228) microsatellite primer sets ware potentially useful for this parentage analysis in the chimeric bull, 78.6% (22/28) of microsatellite present in the chimeric bull were uniquely contributed from the Japanese Black and 21.4% (6/28) from Limousin. This chimeric bull semen was used in producing IVF embryos. The chromosome preparations were made from peripheral lymphocytes. Based on chromosome analysis the Chimera had apparently normal chromosomes (29 acrocentric pairs, one large sub metacentric X chromosome and one small sub metacentric Y chromosome). The proportion of acrosome reacted spermatozoa after 1 h of incubation was higher (p<0.01) with the Chimera than with the Holstein and in Japanese Brown bulls. But did not differ from Japanese Black and Limousin bull sperm. Fertilization rates observed after 5 h of sperm-oocyte incubation with Chimera sperm were higher (p<0.05) than with Japanese Brown and (p<0.01) than with Holstein sperm, but did not differ from Japanese Black and Limousin sperm. The cleavage rates of IVF oocytes inseminated with Chimera sperm were also higher (p<0.001) compared with Holstein, (p<0.01) Japanese Brown and (p<0.05) Limousin, but did not differ from Japanese Black sperm. The blastocyst rates of IVM oocytes inseminated with sperm were higher (p<0.05) than in Limousin, Japanese Brown and Holstein, but did not differ from Japanese Black. Chimeric cattles were produced by aggregation of parthenogenetic (Japanese Brown) and in vitro fertilized (Holstein) bovine embryos at the Yamaguchi Research Station in Japan and by aggregation of parthenogenetic (Red Angus) and in vitro fertilized (Holstein) embryos at the St. Gabriel Research Station in Louisiana. The aggregation rate of the reconstructed demi-embryos cultured in vitro without agar embedding was significantly lower than with agar embedding. The aggregation was also lower when the aggregation resulted from a whole parthenogenetic and IVF-derieved embryos cultured without agar than when cultured with agar. The development rate to blastocysts, however, was not different among the treatment. To verify parthenogenetic and the cells derieved from the male IVF embryos in blastocyst formation, 51 embryos were karyotyped, resulting in 27 embryos having both XX and XY chromosome plates in the same sample, 14 embryos with XY and 10 embryos with XX. The viability and the percentage of zonafree chimeric embryos at 24 h following cryopreservation in EG plus T with 10% PVP were significantly greater than those cryopreserved without PVP. Pregnancies were diagnosed in both stations after the transfer of chimeric blastocysts. Twin male and single chimeric calves were delivered at the Yamaguchi station, with each having both XX and XY chromosomes detected. Three pregnancies resulted from the transfer of 40 chimeric embryos at the Louisiana station. Two pregnancies were Jost prior to 4 months and one phenotypically chimeric viable male born.

• 한국가축번식학회지
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• 제8권2호
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• pp.70-78
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• 1984

In order to investigate the spermiogenesis of Korean cattle spermatozoa, the tissue obtained from the testis and the epididymis of six matured bull were conventionary prepared as thin sections and these sections were observed under a transmission electron microscope. Based on the development of the various permanent organelles and certain transient structures, the spermiogenesis of the Korean cattle could be divided into four main phases: Golgi, cap, acrosome and maturation, and these phases had been further subdivided into a total of nine stages. In the maturation phase, the condensation of the nuclear karyoplasm had progressed from the u, pp.r side to the lower side of the nucleus as the movement of the nuclear ring which would be disa, pp.ared after maturation.

• 한국가축번식학회지
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• 제5권2호
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• pp.43-48
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• 1981

This experiment was carried out with the purpose to get some information about the properties of Simmental bull semen during summer season. The results obtained were summarized as follows: 1. Semen volume per ejaculation, sperm concentration and sperm viability were averaged 5.16ml, 6.6billion and 65%, respectively. 2. Percentage of motile sperm after dilution in skmmilk solution and trisbuffer for 5 days were 34.16% and 35.0%, respectively. 3. Viability of spermatozoa frozen in skimmilk extender and trisbuffer for 5 days were 20. 83% and 25.66%, respectively. 4. Percentage of live sperm, MRT and pH value were 71.8∼72.1%, 8.40∼8.21 minutes and 6.78, respectively. 5. Diluted semen showed strong resistance to the cold shock than that of fresh semen. 6. Rscovery of sperm motility after freezing for 24 hours was relatively weak.

• 한국수정란이식학회지
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• 제27권3호
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• pp.155-162
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• 2012

This study was designed to determine whether low-density lipoporoteins (LDL) extracted from egg yolk in extender improve the function of Korean Jeju Black Bull semen. The semen was cryopreserved with 5% ethylene glycol (EG) or 7% glycerol (G) extenders containing 10% egg yolk (EY), 4% LDL and 5% EY or 8% LDL. Frozen-thawed sperm were evaluated sperm motility, viability, membrane integrity and acrosome integrity. Post-thawed sperm motility has been significantly higher (p<0.05) in 4% LDL + 5% EY ($69.00%{\pm}4.18$; EG and $63.00%{\pm}9.75$; 7% G) than 8% LDL ($57.00%{\pm}5.70$; EG and $52.00%{\pm}4.47$;G). Treatment of 4% LDL + 5% EY-EG ($66.85%{\pm}5.06$) has been significantly improved sperm viability compared to other treatments except 10% EY - EG. Moreover, in membrane integrity, swollen sperm ratio has been only significantly increased (p<0.05) in 4% LDL + 5% EY - EG ($64.65%{\pm}6.10$) among all treatments. In assess to detect acrosome integrity, especially, AR pattern ratio has been significantly decreased (p<0.05) in 4% LDL + 5% EY - EG among all treatments. In sperm viability as time passes, between 4% LDL + 5% EY and 10% EY, there was no significant difference, but 8% LDL was significantly decreased sperm viability in EG (1 and 2 hrs) and G (30 min, 1, 2, 5 and 12 hrs) extender. However, there were no significant differences among all treatments except 8% LDL-G in sperm membrane integrity. 8% LDL-G has been significantly decreased swollen sperm ratio at 5 hrs after thawed. It is concluded from these results that 4% LDL + 5% EY to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Korean Jeju Black bull.

• Asian-Australasian Journal of Animal Sciences
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• 제29권9호
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• pp.1247-1255
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• 2016

Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.