• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.479-483
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• 2011

The objective of this study was to determine the effects of E. coli isolated from porcine semen on sperm viability, motility, and semen pH. Semen samples were prepared using commercial extender, $Seminark^{Pro}$ (Noahbio Tech, Korea) that did not contain antibiotics. And 4 different levels of E. coli were artificially innoculated to semen with following concentrations; 4,000 of sperms with 1 of E. coli (T1), 400 with 1 (T2), 40 with 1 (T3), and 4 with 1 (T4). Semen samples were preserved at $17^{\circ}C$ for 5 days in semen storage box until analyzed by flowcytometer. Aliquots were subjected to measure the sperm viability (Live/$Dead^{(R)}$ stain), motility (mitochondrial function), and semen acidity (pH) from day 0 (day of semen collection) to day 5. Sperm motility and viability were significantly decreased (p<0.05) on day 0 (4 hrs after preservation at $17^{\circ}C$) in T3 and T4 compared to control groups and were significantly decreased (p<0.05) in all groups from day 3. Sample pH was acidic in T3 (6.90~6.86) and T4 (6.86~6.65) from day 3 to day 5 (p<0.05). On the other hand, sample pH was maintained 7.0~7.1 in control, T1, and T2 during the experimental period. Sperm motility and viability were significantly decreased from day 0 to day 5 compared to control in samples contaminated with E. coli above a value of 40:1 ($20{\times}10^6$ sperm cells/ml : $5{\times}10^5$ cfu/ml). Even on day 1 in T4 and on day 3 in T3, semen pH was acidic probably due to the acidification of dead spermatozoa. These results suggest that E. coli contamination has a concentration-dependent detrimental effect on extended porcine semen quality.

• Animal Bioscience
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• v.37 no.2
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• pp.203-209
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• 2024

Objective: This study aimed to assess the impact of the dilution ratio of Tris diluent, storage at 0℃, and long-distance transportation on the spermatozoa of Simmental cattle. It also validated the feasibility of the regional distribution of fresh semen. Methods: In experiment 1, semen was diluted at four dilution ratios (1:6, 1:9, 1:12, and 1:15) to determine the optimal dilution ratio of Tris diluent. In experiment 2, we assessed sperm viability, progressive motility (objectively assessed by computer-assisted sperm analyzer), and acrosome intactness in Tris dilutions kept at constant 0℃ for 1, 3, 6, 9, and 12 days. We compared them to Tianshan livestock dilutions (Commercial diluent). In experiment 3, semen was diluted using Tris diluent, and sperm quality was measured before and after long-distance transport. Artificial insemination of 177 Simmental heifers compared to 156 using Tianshan Livestock dilution. Results: The outcomes demonstrated that 1:9 was the ideal Tris diluent dilution ratio. The sperm viability, Progressive Motility, and acrosome integrity of both Tris and Tianshan dilutions preserved at 0℃ gradually decreased over time. sperm viability was above 50% for both dilutions on d 9, with a flat rate of decline. The decrease in acrosome integrity rate was faster for Tianshan livestock dilutions than for Tris dilutions when stored at 0℃ for 1 to 6 days. There was no significant difference (p>0.05) in sperm viability between semen preserved in Tris diluent after long-distance transportation and semen preserved in resting condition. The conception rates for Tris dilution and Tianshan livestock dilution were 49.15% and 46.15% respectively, with no significant difference (p>0.05). Conclusion: This shows that Tris diluent is a good long-term protectant. It has been observed that fresh semen can be successfully preserved for long-distance transport when stored under 0℃ conditions. Additionally, it is feasible to distribute semen regionally.

• Korean Journal of Veterinary Research
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• v.33 no.2
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• pp.345-350
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• 1993

Multiple ejaculates were collected from four male mongrel dogs. The second fraction and the small volume of third fraction from the ejaculates were divided and treated as follows : control; addition of the egg-yolk Tris extender to the semen at $37^{\circ}C$. group I; Removal of seminal plasma, group II; addition of the glycerolated extender at $4^{\circ}C$, group III Removal of seminal plasma and addition of glycerolated extender at $4^{\circ}C$. The semen cooled to $4^{\circ}C$ was equlibrated for 2hrs and preserved in refrigerator at $4^{\circ}C$. The preserved semen was evaluated for kinetics, morphology, motility and thermoresistance daily for 3 days. 1. The kinectics after preserved days 2 and 3 of group I was significantly higher than that of control(p<0.05). 2. There were no significant difference in abnormal morphology of each group between the periods of storage. 3. The motility after preserved day 1 and days 3 of group I was significantly higher than that of others(p<0.05), and the molity after preserved days 2 of group I and III was signficantly higher than that of others(p<0.05). 4. When the molity of preserved semen was measured during incubation at $37^{\circ}C$, the motility of four groups was declined at similar rates. There was no effect of removal of seminal plasma and glycerol addition on thermoresistance.

• Food Science and Preservation
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• v.14 no.5
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• pp.538-544
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• 2007

This study was conducted to evaluate the antioxidant (ABTS, DPPH radical scavenging and reducing power), nitric oxide (NO)) production and anticancer activities against human cancer cells (HeLa HepG2, HT-29 and MCF-7) for methanol extracts of Glycine Semen Germinatum (Daedoowhangkeun in Korean) fermented with germinated black soybean and some bacteria. Antioxidant activities were increased by increasing the concentration of the extract at dose-dependent manner, Their activities of black soybean were higher than those of yellow soybean. Non fermented sample was slightly higher than Glycine Semen Germinatum fermented with Bacillus subtilis and lactic acid bacteria. In 1 mg/mL of the extract NO production levels were $0.374\;{\mu}M$ for yellow soybean, $0.368\;{\mu}M$ for black soybean, and $0.367\;{\mu}M$ and $0.358\;{\mu}M$ for Glycine Semen Germinatum fermented with B. subtilis and lactic acid bacteria, respectively. Methanol extract of Glycine Semen Germinatum fermented with mixture broth of lactic acid bacteria was shown to be the highest activity for anticancer activities against human cancer cells tested and their activities were exhibited in the order of HeLa > HT-29 > HepG2 > MCF-7 cell.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.2
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• pp.69-75
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• 2021

Cryopreservation is a widely-used efficient means of long-term sperm preservation. However, unlike other types of semen, cryopreserved boar semen has reduced fertility and the efforts continue to optimize post-thawing sperm recovery. In this study, we evaluated the effects of various washing solutions (Hulsen solution, lab-made DPBS and commercial DPBS) on post-thawing porcine sperm kinematics (CASA system), viability (SYBR-14/PI) and acrosome integrity (PSA/FITC). We also examined the effect of washing-centrifugation on frozen-thawed semen kinematics. The results indicate that type of washing solution and post-thawing centrifugation alters parameters linked to sperm quality (total motility, progressive motility, viability and acrosome integrity). Significantly higher (p < 0.05) motility and progressive motility were obtained when cryopreserved semen was processed with Hulsen solution. The post-thaw percentage of live and intact acrosomal sperm was significantly higher in group 1 (Hulsen solution) as compared to other groups. Following thawing-centrifugation, the results showed significantly higher motility and progressive motility in group 1 than other groups. However, the latter two DPBS groups did not differ statistically. Taken together, Frozen-thawed spermatozoa motility, acrosome integrity and viability can be affected by the type of washing solution used. Moreover, centrifugation of frozen-thawed semen has an unfavorable effect on total motility and progressive motility.

• Journal of Embryo Transfer
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• v.18 no.2
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• pp.143-149
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• 2003

This study was carried out to cryopreserve and to investigate characteristics of semen in Hanwoo. Semen was obtained from bulls selected by Daekwanryeong Branch station. Semen was collected each morning of the experiment, placed in water jacketed tubes at 37$^{\circ}C$, and trans-ported to the research laboratory within 10 minutes. Semen was extended with Egg yolk-glycerol extender to contain 50${\times}$10$^{6}$ sperm/ml. Semen was cooled over a 6h period in water jacketed tubes from about 25 to 5$^{\circ}C$, Egg yolk-glycerol extender was added in one step at 5$^{\circ}C$. Semen was aspirated into 0.5ml straws, which were sealed with powder. Egg yolk-glycerol extender, which is used in Hanwoo sperm frozen and stored, semen from 13 Hanwoo bulls collected, the postthawed percentages of motile sperm were 65.7%. In semen characteristics of Hanwoo bulls, number of bulls volume are 5.7 ml and total cell count are 975${\times}$10$^{6}$ m1 ejaculate.

• Animal Bioscience
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• v.35 no.12
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• pp.1827-1838
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• 2022

Objective: The semen quality of stallions including sperm motility is an important target of selection as it has a high level of individual variability. However, effects of the molecular architecture of the genome on the mechanisms of sperm formation and their preservation after thawing have been poorly investigated. Here, we conducted a genome-wide association study (GWAS) for the sperm motility of cryopreserved semen in stallions of various breeds. Methods: Semen samples were collected from the stallions of 23 horse breeds. The following semen characteristics were examined: progressive motility (PM), progressive motility after freezing (FPM), and the difference between PM and FPM. The respective DNA samples from these stallions were genotyped using Axiom Equine Genotyping Array. Results: We performed a GWAS search for single nucleotide polymorphism (SNP) markers and potential genes related to motility properties of frozen-thawed semen in the stallions of various breeds. As a result of the GWAS analysis, two SNP markers, rs1141327473 and rs1149048772, were identified that were associated with preservation of the frozen-thawed stallion sperm motility, the relevant putative candidate genes being NME/NM23 family member 8 (NME8), olfactory receptor family 2 subfamily AP member 1 (OR2AP1), and olfactory receptor family 6 subfamily C member 4 (OR6C4). Potential implications of effects of these genes on sperm motility are herein discussed. Conclusion: The GWAS results enabled us to localize novel SNPs and candidate genes for sperm motility in stallions. Implications of the study for horse breeding and genetics are a better understanding of genomic regions and candidate genes underlying stallion sperm quality, and improvement in horse reproduction and breeding techniques. The identified markers and genes for sperm cryotolerance and the respective genomic regions are promising candidates for further studying the biological processes in the formation and function of the stallion reproductive system.

• Journal of Embryo Transfer
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• v.24 no.1
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• pp.29-32
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• 2009

Prostaglandin $F_2{\alpha}$ ($PGF_2{\alpha}$) can facilitate release of epinephrine from the adrenal gland. The objective was to extend these findings and determine the effects of $PGF_2{\alpha}$ on sexual activity and semen collection training in sexually inexperienced boars. Boars (n=32; $281{\pm}18$ days of age) were moved individually once weekly to a semen collection room equipped with an artificial sow. Before entering the semen collection room, boar received i.m. treatments of $PGF_2{\alpha}$ at doses of 5 (n=8), 10 (n=8), or 20 (n=8), and control boar (n=8) were not treated. Reaction time (elapsed time after entering collection pen until the start of mounting) for boars receiving 5mg ($3.3{\pm}0.9\;min$), 10mg ($3.3{\pm}0.8\;min$) $PGF_2{\alpha}$ was shorter (p<0.05) than for controls ($6.7{\pm}0.9min$). Duration of ejaculation (min) per session was longer (p<0.05) for $PGF_2{\alpha}$ (10 mg, 20 mg)-treated boars ($7.3{\pm}0.7\;min$, $6.9{\pm}0.7\;min$), compared to control ($3.4{\pm}0.8\;min$). The number of training session per boars was less (p=0.056) for $PGF_2{\alpha}$ 10mg-treated boars ($1.0{\pm}0.4$), compared to control ($2.0{\pm}0.4$). Semen characteristic such as volume, concentration, the number of total ejaculated sperm, were similar for $PGF_2{\alpha}$-treated and controls. There was no apparent difference on sperm movement characteristics (Mot: motility, VCL : curve linear velocity, VSL : straight line velocity, VAP : average path velocity, LIN : linearity) after semen preservation by collected with or without $PGF_2{\alpha}$ treatment. In summary, administration of $PGF_2{\alpha}$ in boars increased the sexual activity and facilitated the training boars to mount an artificial sow for semen collection, but did not affect semen characteristic.

• Korean Journal of Poultry Science
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• v.41 no.1
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• pp.21-27
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• 2014

To preserve chicken genetic materials like cryopreserved spermatozoa, various kinds of freezing agents like glycerol, dimethylsuloxide, dimethylformamide or dimethylacetamide have been used for rooster semen preparation. Recently, the usage of N-methylacetamide (MA) for Ogye rooster semen preservation resulted in hatched chicken successfully. In this study, we investigated the effects of 7, 9 and 11% of MA on the viability, fertility and hatchability of frozen-thawed rooster semen using artificial insemination. The results of viability, fertility and hatchability in frozen semen with 7%, 9% or 11% MA were $35.16{\pm}6.12%$, $67.83{\pm}15.3%$ and $66.2{\pm}16.3%$ of motile sperm rate, 21.5%, 34.7% and 25% of fertility rate, and 100%, 89.5% and 87.5% of hatchability rate. The results of control group with frozen semen were 96.0% of fertility rate and 92.2% of hatchability rate. With these results, the concentration range of MA as a freezing agent of rooster semen could be 7~9% of media. The higher concentration of 9 % MA could decrease the fertility rate of thawed semen not the rate of hatchability rate. So the use of MA without affecting fertility rate would be a key point of freezing method of rooster semen for poultry genetic resource preservation.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.9-16
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• 2002

This study was carried out to investigate the effects of extenders such as Beltsville thawing solution(BTS), Modena and Androhep, preservation temperature and period of liquid boar semen on semen characteristics and reproductive performance. Boars were raised at Swine Artificial Insemination Center in National Livestock Research Institute, Sunghwan, Chungnam, Korea. This experiment was carried out from 1995 to 2000. The results obtained were summarized as follows. 1. Sperm motility in the samples with Androhep and BTS reduced from day 5 and in the samples with Modena reduced from day 3 of storage. pH or 3 extenders varied from 6.24 to 7.04 during day 1 to 5 of storage. Farrowing rate of sows inseminated with liquid boar semen offended with BTs, Modena and Androhep extenders did not show any differences until day f after semen collection. Sows inseminated with Androhep extender had better farrowing rates (P＜0.05) than those with Modena extender at day 1 or 5 after semen collection, but farrowing rates after AI using BTS did not differ compared to those Androhep and Modena. Litter size did not show any differences among the three extenders, but Androhep had the decreased litter size from day i of storage. 2. Motility and normal acrosome of the sperm preserved at 5$^{\circ}C$ did not show any differences until day 4 of storage, but those at 17$^{\circ}C$ changed from day 3 and 4, respectively. Farrowing rate of sows artificially inseminated with liquid boa. semen preserved at 17$^{\circ}C$ had higher, than at 5$^{\circ}C$ (p＜0.05), but there was no significant differences in litter size. Farrowing rates and litter size were decreased from day 2 and day 3 of storage at 17$^{\circ}C$, respectively. Farrowing rate of sows inseminated with the preserved semen at 5$^{\circ}C$ did not changed until day 4, but the litter size at 5$^{\circ}C$ was lower than that at 17$^{\circ}C$.