• Asian-Australasian Journal of Animal Sciences
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• v.5 no.3
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• pp.439-447
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• 1992

Eight healthy Holstein bulls, 4-6 years old were used to study the effect of season of the year on the biophysical characteristics of semen. Semen was collected twice a week by AV (artificial vagina) over one-year period. The analyses revealed that all the basic seminal traits studied were differed significantly due to season, except the ejaculate volume and consistency and the percentage of swollen spermatozoa in a hypo-osmotic fructose-citrate solution. Ejaculates collected during hot summer season had significantly lower sperm motility, concentration and total counts, and higher percentage of dead spermatozoa than those collected during winter time. Warm spring had moderate semen quality. The temperature-humidity index was calculated and it was associated (p < 0.01) negatively with the ejaculate pH, sperm concentration and total counts, and positively with the % of dead sperms. Ejaculate volume, percentage of swollen spermatozoa, individual motilities did not correlate significantly with the change in temperature-humidity index values. The total live, motile spermatozoa per ejaculate during both the winter and spring seasons showed significant increase of about 37% and 32% respectively over the summer season. Also, rectal temperatures of the bulls were elevated during the hot summer season, while the values of blood hemoglobin and packed-cell volume were decreased.

• Korean Journal of Animal Reproduction
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• v.3 no.1
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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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• v.13 no.1
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• pp.10-18
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• 2000

This paper describes the magnetic orientation of the intact and demembranated bull sperm treated by DTT or heparin in a 5,400 G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4{^{\circ}C}$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the sperm head area at 1, 3, and 6 days. After measuring the area, each sperm sample was exposed to a 5,400 G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the decondensation of bull sperm nuclei was not induced by the heparin treatment, however, incomplete decondensation was induced by the DTT treatment. During the magnetic orientation, bull sperms treated by DTT or heparin had low percentages of long axis perpendicular to the magnetic lines of force. However, different aspects were obtained for long axis perpendicular orientations following treatment of DTT or heparin. Through the DTT treatment, the decline of long axis perpendicularly oriented percentages was due to the increase of long axis parallel orientation with the head of the flat plane perpendicular to the magnetic lines of force, whereas, using the heparin treatment, the decline of long axis perpendicular orientation was due to the increment of long axis parallel orientation with the head of the flat plane parallel to the magnetic lines of force. Also, percentages of the head of the flat plane perpendicular were decreased by the heparin treatment. These findings suggest that maintaining the structure of protamine in the chromatin is necessary for the sperm head to orient with its flat plane perpendicular, and maintaining the disulfide bond in the chromatin is necessary for the long axis of sperm to orient perpendicularly.

• Korean Journal of Animal Reproduction
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• v.12 no.1
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• pp.24-30
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• 1988

These experiments were carried out to develop new techniques for In Vitro separatin of X-and Y-bearing spermatozoa. The bull semen was applied to the various Gel-Columns filled with swellen Sephadex G-50 Fine and then elutriated wtih Locke solution (elutriation rate; 1ml/3-4min., 1ml/1-2min.). Elutriated solution was fractionated into 1ml by automatic Fraction Collector and spermatozoa included in each fraction were subjected to the estimation of viability and recovery rate, and to B-body test. The results obtained in these experiments were summarized as follows: 1. When the column size and the elutriation rate were adjusted to 15$\times$1.6cm and 1ml/3-4min., respectively, the highest sperm concentration was obtained from the 8th to the 12th fraction. 2. As a trend, the viability of spermatozoa was improved by chromatography, and the degree of improvement ranged 5 to 10 percentage. 3. The average recovery rate of spermatozoa applied to column was 73.2 percentage and ranged 52.6 to 81.3 percentage. 4. The lowest rate of B-body bearing spermatozoa following chromatography was obtained when the column size and the elutriation rate were adjusted to 15$\times$0.8cm and 1ml/1-2min., respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.3
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• pp.335-340
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• 2006

A large number of toxicological substances and pharmacological and physical agents can cause reproductive intervention at the cellular and molecular level. The present study was designed to assess the effect of mercury ($HgCl_2$) at 50 to $550{\mu}M$ concentration ranges, in vitro, on the sperm membrane and DNA integrity, viability, and acrosomal status of normal bull spermatozoa. The samples were processed for sperm analyses using semen-diluting fluid (PBS, pH 7.2). We recorded a sharp increase in the lipid peroxidation/LPO rate; the highest was at $550{\mu}M$ mercury concentration, indicating a deleterious effect of mercury on the sperm membrane intactness. There was also a strong negative correlation between LPO rate and % viable spermatozoa (R = 0.987, p<0.001). Data obtained from a comet assay technique revealed that mercury is capable of inducing DNA breaks in the sperm nuclei. Interestingly, 92% of DNA breaks were double-stranded. The correlation between LPO rate and % DNA breaks was 0.984. Performing the gelatin test indicates that mercury is able to alter the integrity of acrosomal membranes showing an abnormal acrosome reaction. In this regard, a strong link was found between LPO rate and % halos (R = 0.990, p<0.001). Collectively, mercury proved to be a potent oxidant in the category of environmental factors affecting bull spermatozoa. Hence, considering the wide spread use of mercury and its compounds, these metals should be regarded with more concern.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.321-326
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• 2018

In this study, two epididymal spermatozoa recovery methods in relation to sperm number, motility, viability and acrosome reaction were examined. Seven bulls were castrated and 7 testicles with epididymides were transferred to the laboratoy. Epididymis in each bull was randomly used for flushing and mincing methods with semen extender (Optixcell, IMV, France). The recovered spermatozoa with adjusted sperm concentration to $40{\times}10^6cells/mL$ was diluted with optixcell and cryopreserved. In experiment 1, the difference in the total number of spermatozoa using flushing and mincing methods was insignificant (2570.0 and $2505.2{\times}10^6cells/mL$, respectively). For experiment 2, the percentage of motile spermatozoa and motility parameters between flushing and mincing methods were studied through the use of sperm class analyzer after frozen-thawing. The percentage of total motile sperm between flushing and mincing methods was almost the same with $89.5{\pm}12.8$ and $91.4{\pm}7.9%$, respectively. The same is the case with experiment 3 wherein the viability and acrosomal integrity of frozen-thawed epididymal spermatozoa by flushing and mincing was insignificantly different. The results from the study showed that both flushing and mincing methods can be used for epididymal spermatozoa recovery in bull.

• Journal of Embryo Transfer
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• v.26 no.1
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• pp.65-70
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• 2011

This study was conducted to investigate the survival rate of AndroMed and Tris-egg yolk extender for cryopreservation of Korean Native Bull Semen (Chick Cow). Semen was collected from a Korean Native Bull Semen over 3 year's old. The semen was diluted 1:1 by AndroMed and Tris-egg yolk extender. The pellet was diluted to final sperm concentration of $5{\times}10^7$ cell/ml by doubling in every 10 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 1 hrs at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm of liquid nitrogen for 5 minutes. And then the frozen straw was plunged to LN2. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. The survival rates was significantly higher (p<0.05) in Tris-egg yolk extender than AndroMed extender ($89.7{\pm}19.8$ vs. $73.4{\pm}11.2$). However, motility was no significant differences ($78.4{\pm}18.7$ vs. $67.9{\pm}14.6$). Survival rate in time of equilibration between visual and CASA program had higher in 2 h ($86.33{\pm}9.4$ vs. $92.32{\pm}12.4$) than in 5 h ($78.20{\pm}7.8$ vs. $88.28{\pm}13.1$) 15 h ($65.24{\pm}6.6$ vs. $76.48{\pm}17.3$) 20 h ($56.26{\pm}4.6$ vs. $67.73{\pm}18.4$). The development rates to cleavage was higher in Tris-egg yolk extender than AndroMed extender (82.2% vs. 81.7%). Similarly, the development rates to blastocyst was significantly higher (p<0.05) in Tris-egg yolk extender than AndroMed extender (42.3% vs. 29.6%). In conclusion, the obvious impact of this study will be its practical application to improve viability and fertilizing ability of cryopreserved spermatozoa used for in vitro fertilization (IVF) and AI, Which in turn will be beneficial to animal genetic resources conservation.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.197-205
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• 1996

This study was conducted to examine the effect of sustained release recombinant bovine somatotropin(SR-rBST) with different dosage(0, 0.03mg/kg body weight/14 day, 0.06mg/kg body weight/14 day, 0.09mg/kg body weight/14 day) on the economical traits of the semen characteristics and sexual behaviors and blood chemical values with sixteen Holstein dairy young bulls in Dairy Cattle Improvement Center of National Livestock Co-operatives Federation. Sensual testings of collected semens of Holstein young bulls were not different among treatments (p>0.05). Ejaculated semen volumes of control, SR-rBST 0.03mg, 0.06mg and 0.09mg groups were 5.6ml, 5.6ml, 6.0ml and 6.7m, respectively, but the result of SR-rBST 0.09mg group significantly increased semen volume(P<0.05). In sperm concentration, SR-rBST 0.06mg and 0.09mg groups significantly increased the total number of sperms than the other groups(P<0.05). In otherwise, SR-rBST treatments did not affect on pH, osmotic pressure, anti forzen rate, abnormality and motility of collected semen during whole experimental period. Reaction time(RT), sexual aggressiveness(SA) and tactile stimulation(TS) were not different among treatments. The libido scores(LS) of control, 0.03mg, 0.06mg and 0.09mg groups were 68.0, 80.5, 73.9 and 80.8, respectively, and LS were significantly improved by SR-rBST administration(P<0.05). The effect of SR-rBST on scrotal circumference measurement that is important factor to determine the ability of semen production, was not different among treatments. These studies indicates that SR-rBST treatments favourably affect on semen quality and sexual libido in Holstein young bull.

• Journal of Animal Science and Technology
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• v.51 no.2
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• pp.105-110
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• 2009

The Objective of this study was to compare performances of proven and young holstein bulls bred in Korea. Proven bulls are categorized into the imported and the korean ones. Data from 148,329 heads of daughters of 1,128 bulls from 1990 to 2004 were used in this study. Proven bulls showed higher milk yield than young bulls in same year. Young bulls, however, always yielded more milk than korean bulls when proven bulls were categorized into the imported and the korean ones. Hence, it was proven that dairy bull selection program had properly been functioned in Korea. Selected bulls, which were korean proven bulls and young bulls, yielded higher milk fat than imported bulls as the selection was weighted on the yield of the milk fat. This comparison was based on the performances of daughters without the consideration of the semen price. Semen price of the imported proven bulls were higher than the korean proven bulls and the semen of young bulls was free. Hence, the performances of korean bulls with the consideration of the preferential effect would be much higher than others, and further studies are necessary.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.2
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• pp.91-98
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• 2021

The objective of this study was to establish a selection process for high quality sperm in bovine semen using sperm separation by magnetic activation (MACS). For this, semen from 21 Nellore bulls was collected using an artificial vagina. To guarantee the presence of pathologies in the ejaculate, animals previously declassified in four consecutive spermiogram were used. Semen was analyzed in five statuses: (1) fresh semen (fresh); (2) density gradient centrifugation (DGC), percoll column; (3) non-apoptotic fraction after separation by MACS (MAC); (4) apoptotic fraction from the separation (MACPOOR); and (5) MAC followed by DGC (MACDGC). Using a computerized analysis system (CASA), motility was measured. The sperm morphology was evaluated by phase contrast, and the supravital test was completed with eosin/nigrosin staining. For DGC, 20 × 10⁶ cells were used in a gradient of 90% and 45% percoll. MACS used 10 × 10⁶ cells with 20 μL of nanoparticles attached to annexin V, and filtered through the MiniMACS magnetic separation column. Membrane integrity was assessed with SYBR-14/IP and mitochondrial potential with JC-1 by flow cytometry. Processing sperm by MACDGC, was more effective in obtaining samples with high quality sperm, verified by the total of abnormalities in the samples: 35.04 ± 2.29%, 21.50 ± 1.47%, 17.30 ± 1.10%, 30.68 ± 1.94% and 10.50 ± 1.46%, respectively for fresh, DGC, MAC, MACPOOR, and MACDGC. The subpopulation of non-apoptotic sperm had a high number of live cells (82.65%), membrane integrity (56.60%) and mitochondrial potential (83.98%) (p < 0.05). These findings suggest that this nanotechnological method, that uses nanoparticles, is efficient in the production of high-quality semen samples for assisted reproduction procedures in cattle.