• 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.

• Korean Journal of Animal Reproduction
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• v.7 no.2
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• pp.57-71
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• 1983

1. Diluted chicken semen can be preserved at 2 to 5$^{\circ}C$ for 24 to 48 hr with resultant fertility of greater than 90% of that of fresh semen. Turkey semen can be preserved at 10 to 15$^{\circ}C$ for 6 to 24 hr and provide economical fertility. 2. Frozen chicken semen has given variable results; a 21 to 93% fertility ranges as compared to 92 to 94% expected with fresh semen. Highest fertility levels obtained with frozen turkey semen intravaginally inseminated have been 61 and 63% using DMSO and glycerol, respectively, as cryoprotectants. 3. The use of glycerol as a cryoprotectant reauires that its concentration in semen be reduced to less than 2% either by dialysis or centrifugation after thawing and before intravaginal insemination if optimal fertility is to be obtained. 4. The temperature at which cryoprotectants are added to semen and the time allowed for equilibration are important for subsequent fertility pre- and post-freezing. 5. The type of container used for packaging the semen, freeze or cooling rates, thaw rates and level of cryoprotectant all interact in affecting cell survival. 6. Plastic freeze straws as a packaging device for semen offers the following advantages: easy to handle, require minimal storage space, offer a wide range of freeze and thaw rates, and insemination can be made directly from them upon thawing. 7. Controlled slow cooling rates of 1 to 8$^{\circ}C$/min have thus far provided the best results for cooling chicken semen throught the transition phase change (liquid to solid) or critical temperature range of ＋5 to -20 or -35$^{\circ}C$. 8. Highest fertilities have been achieved with frozen chicken semen where a slow thaw rate (2。 to 5$^{\circ}C$) has been used regardless of the freeze rate. 9. To maintain a constant high level of fertility throughout a breeding season with frozen semen, a higher absolute number of spermatozoa must be inseminated (2 to 3 times as many) as compared to fresh semen since a, pp.oximately 50% are destroyed during processing and freezing. 10. The quality of semen may vary with season and age of the male. Such changes in sperm quality could be accentuated by storage effects. Thus, the correct number of spermatozoa may very well vary during the course of a breeding period. 11. As to time of insemination, it is best to avoid inseminating chicken hens within 1-2 hr after or 3-5 hr before oviposition; and turkey hens during or 7-10 hr before oviposition. 12. The physiological receptiveness of the oviduct at the time of insemination is a very important biological factor influencing fertility levels throughout the breeding season.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.221.1-221.1
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• 2003

Armeniacae Semen is a seed of Prunus armeniaca Linne var. ansu Maximowicz, which belongs to Rosaceae family. It contains amygdalin and fatty oil and is widely used to treat asthma, dysponea and edema. It was reported that D-amygdalin in Armeniacae Semen undergoes hydrolytic reaction by emulsin when using water, and esecially it is almost decomposed when extracting from powder type. we set up a condition where we can achieve the maximun extraction yield through the study of the methods to rstrain emulsin from causing hydrolysis of D-amygdalin in Armeniacae Semen in the aqueous solution and to prevent D-amygdalin from being converted into neoamygdalin. (omitted)

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.239-243
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• 2008

This study was conducted to investigate the effects of type of the sugar supplemented to the extender on the vigor, viability and intact acrosomal rates of frozen-thawed dog spermatozoa. The ejaculated semen was diluted with TRIS-citric acid extender containing 200mM TRIS, 73mM citric acid, 6% (v/v) glycerol, 20% (v/v) egg yolk, 1% (v/v) antibiotics (streptomycin/penicillin), 44 mM sugar, which was either glucose, fructose or glucose-fructose combination, and distilled water to make the final volume of 100ml. Extended semen samples were cooled at $4^{\circ}C$ for an hour, packaged in 0.25ml straws, equilibrated for 10 minutes in liquid nitrogen vapor, and frozen in liquid nitrogen. Samples were thawed by placing straws into $37^{\circ}C$ water for 120 seconds. After thawing, vigor, viability and intact acrosomal rates of frozen-thawed semen were compared according to type of sugar. No significant differences were observed between glucose and fructose groups. In addition, combination of the 2 sugars also did not show any significant differences in the vigor, viability and intact acrosomal rates. In conclusion, glucose and fructose were equally efficient as sugar supplements for freezing extender.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.71-79
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• 1992

In vitro fertilization and embryo transfer (IVF & ET) is widely used for the males with subnormal or abnormal semen quality, as this was recommended in view of the relatively small numbers of spermatozoa required for fertilization and subsequent pregnancies could be obtained. The aim of this study is to know how the various functional parameters of spermatozoa in semen analysis affect the outcome IVF. This study was carried out between 1988-1989, with male factor patients selected on the basis of the semen quality. The selection criteria was based upon the mean values of concentration,% motility and % normal morphology from at least two semen analysis. There is a significant decrease in the fertilization and embryo transfer rates in the study group compared with control group (35.9% vs. 68% and 48.6% vs. 85.5% respectively), however, there was no significant difference in the pregnancy or delivery rates (19.6% vs. 21.4% and 60.0% vs. 62.5% respectively) per embryo transfer cycles. Fertilization rate is variously affected by the type and degree of sperm defect. No pregnancy was occurred in triple defect group and asthenoteratospermia group. There is no significant increase in the abortion rate in the male factor group. Improvement have to be made with the fertilization rate, as the pregnancy rate per OPU cycle in male factor group is still lower than that of normal group (9.5% vs. 18.3%). In conclusion, IVF can be used as a treatment for male factor infertility and the preparation of the semen sample can be modified to improve sperm recovery and obtain fertilization from abnormal semen samples.

• Korean Journal of Animal Reproduction
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• v.12 no.2
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• pp.112-119
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• 1988

These studies were conducted to investigate the effects of culture temperature and time on the in vitro maturation and semen type and media on the in vitro fertilization of bovine follicular oocytes, and to asses in vitro fertilization rate of oocytes cultured by extraffollicular method following fertilization in vitro, or transfer into the pseudopregnant rabbit oviduct or uterus. The bovine oocytes recovered from follicles were cultrued for 18 hrs or 72hrs at 38$^{\circ}C$ with 5% CO2 in moist air. Flesh-diluted(2 folds) and frozen-thawed semen in 0.5ml straw from a fertile bull were used. In order to obtain capacitation of spermatozoa were treated with bovine follicular fluids(BFF) and Inophore A(IA). The results obtained were summarized as follows: 1. The oocytes were classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 62.0%, 12.0%, 17.2%, 5.9% and 3.0% of the total oocytes harvested, respectively. 2. The oocytes matured to metaphase II were significantly increased between 24-48hrs of incubation and at 37-39$^{\circ}C$ with 5% CO2 in moist air. 3. The in vitro fertilization rate following transferred into rabbit oviduct or uterus with bull semen and in vitro matured oocytes were higher ligation than non-ligation of oviduct or uterus. 4. The in vitro fertilization rate of oocytes matured in vitro were higher neat than frozen semen and treatment of IA than BFF on the capacitation of spermatozoa. 5. The effects of semen types and media on in vitro fertilization of oocytes matured in vitro were higher fertilization rate of neat than friozen semen, and media was not significant.

• The Journal of Internal Korean Medicine
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• v.16 no.1
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• pp.40-61
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• 1995

This study was performed to investigate concept, systoms, causes of disease, pathogenic mechanisms, therapies and precriptions about senile constipation through the successive medical literature, recent chinese medical literature and chinese medical joumals. Senile constipation seems to be applicable to dryness syndrom and constipation of insufficiency type, have something to do with kidney(the most), lung, spleen and large intestine. The most principal cause of disease is yin-fluid, the rest deficiency of qi, insufficiency of yang, stagnation of qi and retention of fever etc. There are enriching the blood and moistening dryness in principal therapy, the rest are invigorating qi and loosing the bowel, warming and invigorating the spleen and kidney, regulating the flow of qi and promoting the stagnancy of qi and expelling the pathogenic heat etc. In prescriptions there are Yunjangtang, Jengaektang, Hwanggitang, Jechunjeon, Yukmatang and Majainhwan as the causes of disease, meanwhile are Yungjang-tang, Jechunjeon and Majainhwan in the vulgaris prescriptions. And in medical herbs there are nourishing yin medicines as Rhizoma rehmanniac, Radix ophiopogonis and Radix scrophulariae etc., invigorating qi medicines as Radix astragali, Radix codonopsitis and Radix polygoni multiflori etc, invigorating yang medicines as Caulis cistanchis and Semen psoraleae etc., promoting qi circulating medicines as Radix saussurea, Lignum aquilariae and Radix linderae etc., and reducing fever and therapeutic method to keep the adverse qi flowing downward medicines as Semen cannabis, Rhizoma rhei, Fructus immaturus ponciri, and Cortex magnoliae etc.. Meantime Rhizoma rehmanniae, Radix ophiopogonis, Caulis cistanchis, Radix angelicae gigantis, Semen cannabis, Semen biotae, Semen pruni japonicae and Semen persicae in principal herb-medicines. In clinical reports the process of disease was between 10 to 20 years, the evacuation cycle between 4 to 7 days, generally possessed chronic diseases as hypertension, diabetes, arteriosclerosis and cerebro- vascular disorders etc. and the efficiency rate was more than 90%. The senile constipation is occured in succession or promoted by chronic diseases as obesity, hypertension, diabetes, arteriosclerosis. hrperlipemia, cerebro- vascular disorders etc., so diet-regulating, adequate exercise, proper evacuation-habit and psychologic rest etc. are important more than medicine-treatments.

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.27-34
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• 2006

This study was conducted to compare the reproduction ability of the wild type boar and recombinant human erythropoietin (hEPO) transgenic boar semen. Ejaculated boar semen was analyzed by flow cytometry, Elisa and IVF methods. In experiment 1, flow cytometric analysis showed that the live sperm ratio of transgenic boar sperm significantly lower (P<0.05) than that of wild type boar after incubation at 20, 22, 24 and 26 hr. In experiment 2, the presence and levels of various cytokines (IL-6, IL-10 and $TNF-{\alpha}$) to related animal reproduction in the seminal and blood plasma were examined using specific enzyme immunoassay. There was no significant difference between both groups. In experiment 3, the fertilizing capacity and developmental ability of both boar sperm were compared. The transgenic boar sperm had a significantly low capacity of penetration, sperm-zona binding, embryo development, and blastocyst formation compared to wild type sperm (P<0.05). These results suggest that transgenic boar sperm harboring hEPO gene has low sperm viability than wild type boar, and it is a reason to decrease of fertility and litter size.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.3 s.47
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• pp.389-392
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• 2004

We screened several materials to stimulate IGF-1 promoter activity using luciferase reporter assay and found that Amomi Semen extract (ASE) among them is the most powerful stimulator We also studied about the anti-wrinkle effect of ethanolic extract of Amoni Semen in vitro and in vivo. Semi-quantitative RT-PCR showed that the extract elevated the presence level of IGF-1 mRNA. And $[^3H]$ proline incorporation and semi-quantitative RT-PCR showed that the extract increased the expression of type-I collagen compared with vehicle in vitro and in vivo, respectively. Significant inhibition of MMP-1 expression was determined by ELISA and Western blot. Finally, topical treatment of the extract on hairless mouse's dorsal skin expanded the volume of collagen and dermal thickness. These results suggest that Amomi Semen may be a good candidate for improving extracellular matrix through the increase of collagen expression and inhibition of MMP-1 expression. Moreover, this study enables us to guess that IGF-1 stimulated by the extract may be involved in the mechanism of anti-wrinkle effect of it.

• Korean Journal of Poultry Science
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• v.48 no.4
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• pp.185-191
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• 2021

Chicken spermatozoa have the ability to survive in low-temperature environments; however, the effects of low temperature on sperm motility and acrosome damage have not been studied in detail. The present study investigated semen longevity following dilution of rooster semen with Beltsville Poultry Semen Extender (BPSE) and Lake extender in preservation vessels (1.5 mL e-tube and 0.5 mL straw). Spermatozoa motility in the closed-type vessel (0.5 mL straw) was higher than that in the 1.5 mL e-tube on day 3 of preservation (68.6±3.1% vs. 22.1±5.7%). The motility of rooster semen diluted with BPSE in 0.5 mL straw was also higher than that of the Lake extender on day 3 of preservation (57.7±5.6% vs. 37.7±5.4%). Furthermore, acrosome intactness was higher in 0.5 mL straw than in the 1.5 mL e-tube, and the rate of acrosome cap damage increased with preservation days. The present study demonstrates that a closed 0.5-mL straw vessel could be used for low-temperature semen preservation, with an increased motility rate and acrosome integrity in fresh rooster semen.