• Journal of Embryo Transfer
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• v.21 no.3
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• pp.263-271
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• 2006

The purpose of this study was undertaken to compare ability of frozen-thawed sperm characteristics between two strains (miniature pig and Duroc). The semen was collected by gloved-hand method into a pre-warmed ($37^{\circ}C$) thermos bottle. The semen was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. The frozen-semen straw were thawed at 20, 37 and $50^{\circ}C$ for 1 min, 45 sec and 10 sec within water-bath. The semen sample were evaluated at 0, 3, 6, 9, and 12 h after incubation at $37^{\circ}C$ for analysis of sperm ability. Abnormality of spermatozoa in miniature pig was significantly (p<0.05) higher than that in Duroc at 0, 9 and 12 h of post-thawing incubation after frozen-thawing. The percentage of F-patterned spermatozoa in miniature pig was significantly (p<0.05) lower, while the percentage of AR (acrosome reacted spermatozoa) pattern was higher in the miniature than in the Duroc. On the other hand, there was no significant difference in the viability of spermatozoa thawed at different temperature ($20^{\circ}C\;and\;37^{\circ}C$) between two species, but the viability in miniature pig was higher (p<0.05) than in Duroc when sperm was thawed at $50^{\circ}C$. In conclusion, this study suggest that suitable freezing method for miniature pig semen is required for increasing post-thawing viability and fertilization capacity.

• Journal of Environmental Science International
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• v.6 no.1
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• pp.45-52
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• 1997

Methyl methanesulfonate (MMS) was fed to Drosophila melnogaster in order to investigate its toxic capability at developmental and adult stages, and the hereditary effect of toxicity and the potency for induction of sex-linked lethal mutation during the slyer-matogenesis by the means of an attached-X method. In the control group, the egg to adult viability of D. melnogaster was 95.2%, while 3. 5mM and 5.0mM treated groups were 90.0% and 84.1%, respectively. In the case of their progenies (Fl), the viability was 96.9% in the control group, while 3.5mM and 5.0mM treated groups were 54.5% and 1.6%, respectively. Therefore, these differences between two generations show significant physiological toxic effects in the next generation. In the parental generation, the developmental time was calculated 11.05 days in the control group, 12.43 days In 3.5%mM treated group, and 13.23 days in 5.0mM. In the case of Fl it was estimated 10.35 days in the control group, and 11.43 days In 3.5mM treated group. Compared with the control groups In two generations, the developmental time generally delayed as the dose of MMS increased. As to the sex-ratio, there was no differences between the control and MMS treated groups. The toxic values of adult stage showed which increased the frequency of mortality with MMS concentrations. The mortality at 120hr In the control group was 1.67% and it in 0.5mM MMS treated group 3.33%. In 2.5mM MMS treated group, it was 33.3% at 72hr, and it 95% at 120hr The increase of the morality was shown from 72hr in 4.0mM treated group which was 100% at 96hr. There was the concentration-dependent induction of sex-linked lethal mutation during the spermatogenesis by means of an attached-X method, MMS had more pronounced effect in sperm and spermaid stages in D. melnogaster.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.81-87
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• 2007

The purpose of the present study was to determine the preventive effects of the two antioxidant vitamin E and catechin on DEHP-induced disturbance of spermatogenesis in male rats. Rats at 4 weeks of age were randomly allocated into five groups with 20 animals per group. The first group was not any administrated as control. The second group was administrated DEHP (2 g/kg) daily for 14 days. The third group was administrated vitamin E (500 IU/kg) following DEHP treatment by the same method (daily for 14 days). The fourth group was administrated catechin (200 mg/kg) following DEHP treatment by the same method. The fifth group was co-administrated vitamin E (500 IU/kg) and catechin (200 mg/kg) following DEHP treatment by the same method. In order to determine the preventive effects, we examined pathological changes of testis with apoptotic index, and characteristics of sperm with computer assisted sperm analysis (CASA). Vitamin E and catechin supplementation were significantly prevented the testicular atrophy, apoptosis of germ cells in the seminiferous tubules and abnormal rate of sperm. Moreover, sperm concentration, viability and motility was significantly recovered in groups of alone and along with vitamin E and catechin. The results suggest that preventive effects of alone and along administration of vitamin E and catechin on DEHP-induced testicular atrophy damages have been demonstrated.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.155-162
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• 2005

The present study evaluated whether an exogenous antioxidants, taurine and $\alpha$-tocopherol, could, when added to the freezing extender, improve the post-thaw sperm characteristics, function, the level of reactive oxygen species (ROS) generation, and the level of lipid peroxidation (LPO) in frozen-thawed porcine semen. CASA (computer-aided sperm analysis), HOST (hypoos-motic swelling test), chemiluminescence using luminol and lucigenin and the detection of malondialdehyde for LPO was performed in frozen-thawed porcine sper-matozoa. The results obtained in these studies are as follows. While no beneficial effects of taurine and $\alpha$-tocopherol supplementation were visible in motility, viability, acrosome reaction, tail swelling patterns, and the generation of $O^{2-}$ of frozen-thawed porcine sper-matozoa, $H_{2}O_{2}$ was decreased by all treatments except taurine 50mM treatment. In conclusion the taurine and $\alpha$-tocopherol treatments during freezing reduced generation of reactive oxygen species and production of malondialdehyde in frozen-thawed porcine semen, and the ROS savangers may minimize various damages of spermatozoa during freezing.

• Korean Journal of Veterinary Service
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• v.40 no.3
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• pp.201-208
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• 2017

BVDV causes significant infections in ruminants, resulting in reproductive disorders, diarrhea, reduced milk production and enormous damage to farms. In particular, identification and culling of persistent infectious calf is an important task to eliminate infectious nidus in cattle households. However, studies on physiological characteristics of PI bull are still insufficient to understand reproductive effects of BVDV. In this study, one PI bull was confirmed in herd and complete blood analysis was performed. The lymphocyte count of PI at age 4 was below the normal range and the number of WBCs was also in the lower level of normal range in blood. The sperm number produced by PI male becomes lower and the viability of fresh sperm comes to poor with ages (P<0.05). The sperm abnormality was also increased, especially in nuclear vacuoles of head and droplets of midpeace (P<0.05). The PI male becomes infertile due to poor semen quality at age 4. With these results, we concluded that BVDV in PI bull cause decreased sperm cell and abnormality in semen so causes infertility. However, it appears that BVDV could not be transmitted by indirect contact of PI bull, because there was no evidence of BVDV infection in the herd, when regular vaccination program was applied.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.233-242
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• 1993

The effects of in vitro maturation and sperm treatment condition on the in vitro fertilization (IVF) and developmental capacity of bovine oocytes were investigated and the development of embryos was compared under the 2 different co-culture system, with GC or BOEC. The cultured embryo to 16 cell or morula wre transferred into recipients or frozen by 2 different freezing method. The results obtained were summarized as follows; 1. In vitro maturation rates of vovine follicular oocytes cultrued in TCM199 with 10% FCS or ECS were 64.0% and 72.7%, but the case of addition of 10% FCS or ECS to TCM199 co-cultured with granulosa cells were 81.3% and 84.0%, respectively. IVM rate of three TCM199 added to granulosa cells was higher than that of media without granulosa cells. 2. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC and then fertilized in vitro by sperm treated with caffeine, embryo developments of bovine oocytes co-cultured with BOEC were 38.4% and 51.4%, respectively. But those of bovine oocytes co-cultured with GC were 52.2% by sperm treated with caffeine-heparin. 3. Cleavage rates of bovine oocytes cultured with 10% FCS alone and fertilized in vitro by sperm treated with caffeine-heparin was 33.0%. 4. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC, embryo developments of bovine ooctyes co-cultured with BOEC of GC were 46.0% and 50.2%, respectively. 5. When bovine follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developed co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developments of the bovine oocyte co-cultured with BOEC and GC were 41.8% and 60.1%. But with FCS 10% those of the bovine oocytes co-cultured with BOEC and GC were 42.0% and 48.4%, respectively. 7. When Holstein's follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments fo the bovine oocytes co-cultured with BOEC and GC were 50.0% and 57.7%, but with ECS 10% those of the bovine oocytes co-cultured with BOEC and GC were 52.2% and 56.5%, respectively. 8. The viability of frozen-thawed embryos ranged from 60~80% and those of frozen-thawed embryos from vitrification was lower than that from conventional metiod. 9. The selected fresh embryos were transferred nonsurgically to 7 recipients but did not result in pregnancy.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1160-1168
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• 2018

Objective: This study was conducted to compare morphological defects, viability, motility (MOT), fertility (F), and hatchability (H) in four Korean native chicken breeds (KNCBs), and to evaluate whether defective segments of spermatozoa are associated with MOT, F, and H. Methods: Four KNCBs, including Korean Ogye (KO), Hwangbong (HB), Hyunin Black (HH), and Hoengseong Yakdak (HY) were used. White Leghorn (WL) was used as a control. Nine cocks from each breed were randomly assigned into three groups. Semen was collected by abdominal massage method. Eosin-nigrosin staining method was used to identify live-dead spermatozoa. Different segments and specific morphological defects of spermatozoa were identified using 4', 6-diamidino-2-phenylidole and MitoTracker Red CMXRos. F and H rates were evaluated following artificial insemination (AI). Results: KO had the highest MOT rate compared to HY. Viable normal sperm rates of KO and HH were high and comparable with WL. HY spermatozoa had the highest viable abnormal sperm (VAS) or morphological defect rate followed by HB. Likewise, HB spermatozoa had the highest dead sperm (dead) rate compared to KO, HY, and WL. Bent, coiled, detached, broken, and knotted were common identified specific morphological defects for all breeds. Most morphological defects were at the head and tail in all breeds. VAS showed strong negative correlation with MOT (r = -0.697) and F (r = -0.609). Similarly, defective tail was negatively correlated with MOT (r = -0.587), F (r = -0.797), and H (r = -0.448). The F and H rates of KO and WL were comparable. Conclusion: These data indicate that most identified specific morphological defects are at the head and tail. VAS and defective tail were associated with poor motility, F, and H. KNCBs showed more morphological defects than WL. Finally, these results will facilitate successful AI and semen cryopreservation.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.225-230
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• 1992

Many transgenic mice expressing human growth hormone gene were infertile. To investigate the infertility of these transfenic mice, it was looked into the estrus cycle and sexual behaviour and also tested through in vitro fertilization whether the germ cells of these mice normal or not. The infertile female transgenic mice were mated to the fertile males of ICR strain, but in almost all of them the vaginal plugs were not detected and their estrus cycles by vaginal smear were almost irregular which kept up estrus or diestrus stage. Many male transgenic mice did not have the ability of sexual behaviour. Therefore the viability of germ cells in infertile male transgenic mice was investigated by in vitro fertilization, but the sperm were normally fertilized with the eggs and the transgene of parent was passed on to the progeny. These results consequently suggest that the infertility of transgenic mice experssing human growth hormone gene may be due to the physiological activity of human growth hormone, not germ cells.

• Korean Journal of Oriental Medicine
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• v.12 no.1
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• pp.91-102
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• 2006

We treated KunYangDan (KYK) in SD rats in order toexamine the protective effect against cell damage induced by diethylhexyl phthalate (DEHP). KYD reduced DEHP toxicity by increasing the function of immune cell numbers weight increase of spleen.red blood cells,HB,and HCT content,sperm number and mobility,resulting in improving reproductive function by judging from the recovery of testosterone content. Interestingly the hormone change of testosterone by KYD significantly recovered the decreased its leve. Taken together,these results suggest that KYD specifically affect the reproductive function induced by DEHP,an endocrine disruptor.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.166-172
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• 2019

Objective: In vitro maturation (IVM) of immature oocytes can be useful for some infertile patients. In IVM programs, the rates of embryo formation and pregnancy are low. Therefore, it is essential to recognize the main factors involved in regulating oocyte maturation in vitro. The purpose of this study was to investigate the effects of growth differentiation factor 9 (GDF9) and cumulus cell (CC) supplementation in IVM medium on the rates of embryo formation and viability of human blastocysts. Methods: A total of 80 germinal vesicle oocytes from stimulated cycles underwent an IVM program. The oocytes were divided into four groups, where group I consisted of IVM media only and served as the control, group II consisted of IVM+CCs, group III consisted of IVM+GDF9 (200 ng/mL), and group IV consisted of IVM+CCs+GDF9 (200 ng/mL). Intracytoplasmic sperm injection was performed on the IVM oocytes, and the cleavage embryos that were generated were vitrified. Following thawing, the embryos were cultured for 3 additional days, and the viability rates of the developed blastocysts were determined. Results: The maturation rate of the oocytes did not differ significantly across the four groups. The fertilization rate in group II was significantly higher than that in the control group (76.5% vs. 46.2%). Embryo formation was significantly more frequent in all experimental groups than in the control group, while blastocyst formation did not show significant differences in the three experimental groups compared to the control. The mean viability rates in groups II, III, and IV were 58.16%, 55.91%, and 55.95%, respectively, versus 37.78% in the control group (p< 0.05). Conclusion: Supplementation of IVM culture media with GDF9 and CCs enhanced the fertilization, embryo formation, and viability rates of blastocysts generated from vitrified cleavage embryos.