• Journal of Animal Science and Technology
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• v.48 no.6
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• pp.785-796
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• 2006

This study was designed to evaluate the changes in sperm motility, viability, HOST(hypo-osmotic swelling test), IVP(in vitro penetration), SCSA(sperm chromatin structure assay) during storage of liquid semen collected from boars with different farrowing rates using AI, and to find the relationship between boar fertility through AI and sperm diagnostic parameters during semen storage. The results of HOST were significantly decreased according to the increasing of in semen storage days and the results of IVP were significantly decreased at 3 days of semen storage (P<0.05). The %Red was significantly different among the >80%, 70󰠏80% and <70% farrowing rate group at semen storage day 6(P<0.05). The correlation coefficients between the %Red and farrowing rate were increased according to the semen storage. In conclusion, these results suggest that the sperm parameters evaluated in these studies may be useful indicators to predict the fertility of AI and evaluate the semen quality in boars.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.1
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• pp.63-71
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• 1991

To establish the normal fertile range in the results of the sperm zona-free hamster ova penetration assay (SPA) in Korean male, SPA using the low temperature ($4^{\circ}C$) capacitation in TEST-yolk buffer (TYB) was performed in 67 fertile and 26 infertile men. Sperm parameters in routine semen analysis were also checked and compared with the results of SPA. Sperm concentration, motility and motility index (MI) were significantly higher in fertile group compared with infertile group: $96.0{\pm}46.6$ vs $43.6{\pm}31.9{\times}10^6/ml$, $65.5{\pm}14.8%$ vs $45.8{\pm}23.6%$ and $46.31{\pm}13.29$ vs 27.40{\pm}17.98$, respectively. In fertile group, the hamster ova penetration rate (PR) was $98.5{\pm}5.0%$ (80%-100%), and the penetration index (mean penetrations per ovum, PI) was $9.59{\pm}6.35$(3.1-29.0). All the fertile men showed PI>3.0. In infertile group, PR was $24.6{\pm}24.8%$ (0%-70%), and PI was $0.40{\pm}0.42$ (0-1.3). Both PR and PI were significantly lower in infertile group. There was a significant correlation beween PI and sperm motility or MI, respectively, in fertile group whereas there was no correlation in infertile group. These data suggest that SPA using the low temperature capacitation in TYB can be a valuable diagnostic tool for the assessment of male fertility in vitro and provide an important supplement to the traditional tests of sperm quality.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.9
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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.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.168-179
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• 2020

Objective: This study investigated testicular oxidative stress status and physiomorphological function in Wistar rats fed with yaji and cadmium chloride (CdCl₂). Methods: Sixty male albino Wistar rats (12 per group) were randomly assigned to five groups: group I (control), group II (300 mg/kg.bw of yaji), group III (500 mg/kg.bw of yaji), group IV (2.5 mg/kg.bw of CdCl₂), and group V (2.5 mg/kg.bw of yaji+4 mg/kg.bw omega-3). Each group was evenly subdivided into two subgroups and treatment was administered for 14 days and 42 days, respectively. Semen quality (sperm count, progressive motility, normal morphology, and gonadosomatic index), hormones (testosterone, follicle-stimulating hormone, and luteinizing hormone), testicular oxidative stress markers (superoxide dismutase, catalase, glutathione peroxidase, and malonaldehyde) and testicular histomorphological features were examined. Results: Yaji caused significant (p< 0.05) dose- and duration-dependent reductions in semen quality, the gonadosomatic index, testosterone, follicle-stimulating hormone, and luteinizing hormone. Yaji also caused significant (p< 0.05) dose- and duration-dependent decreases in superoxide dismutase, catalase, and glutathione peroxidase activity, as well as increased testicular malonaldehyde levels. Yaji induced distortions in the testicular histological architecture. CdCl₂ damaged testicular function by significantly (p< 0.05) reducing semen quality, reproductive hormone levels, and oxidative stress markers in albino Wistar rats. CdCl₂ also altered the histology of the testis. Conclusion: This study shows that yaji sauce has similar anti-fertility effects to those of CdCl₂, as it adversely interferes with male reproduction by impairing oxidative stress markers and the function and morphological features of the testis.

• Animal Bioscience
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• v.34 no.5
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• pp.844-854
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• 2021

Objective: The potential of extra virgin olive oil (EVOO), betaine (BET), and ginger (GIN), as natural antioxidants, in reducing negative effects of heat stress on physiological responses, antioxidant capacity, semen quality and fertility of bucks under heat stress were investigated. Methods: Forty adult Animal Production Research Institute line rabbit bucks were distributed randomly into four experimental treatments of ten rabbits each. The first treatment was fed the commercial pellet diet (CPD) without supplementation and served as a control. The other three treatments were fed CPD supplemented with EVOO (300 mg), BET (1,000 mg), and GIN (200 mg) per kg diet for 3 consecutive months during the summer season. Results: Supplementation of EVOO, BET, or GIN improved (p<0.05) the sexual desire, progressive motility, vitality, intact acrosome and membrane integrity, sperm cell concentration, sperm outputs and fertility. Seminal plasma total proteins, globulin, total antioxidant capacity, glutathione and glutathione S-transferase, and initial fructose increased (p<0.05), while total lipids, aspartate and alanine aminotransferases and malondialdehyde decreased (p<0.05) compared with the control. In comparing the natural antioxidants treatments, GIN evoked the largest improvement. Conclusion: The inclusion of GIN (200 mg/kg diet) appeared to improve the sexual desire, semen quality and oxidative stress of bucks. This may be a beneficial supplement for the management of rabbit bucks used in natural mating or artificial insemination.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.2
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• pp.135-141
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• 2022

Objective: This study investigated the impact of two stimulation protocols using highly purified human menopausal gonadotropin (HP-hMG) on the endocrine profile, follicular fluid soluble Fas levels, and outcomes of intracytoplasmic sperm injection (ICSI) cycles. Methods: This prospective clinical trial included 100 normal-responder women undergoing ovarian stimulation for ICSI; 55 patients received concomitant follicle-stimulating hormone (FSH) plus HP-hMG from the start of stimulation, while 45 patients received FSH followed by HP-hMG during mid/late follicular stimulation. The primary outcome was the number of top-quality embryos. The secondary outcomes were the number and percentage of metaphase II (MII) oocytes and the clinical pregnancy rate. Results: The number of MII oocytes was significantly higher in the concomitant protocol (median, 13.0; interquartile range [IQR], 8.5-18.0 vs. 9.0 [8.0-13.0] in the consecutive protocol; p=0.009); however, the percentage of MII oocytes and the fertilization rate were significantly higher in the consecutive protocol (median, 90.91; IQR, 80.0-100.0 vs. 83.33 [75.0-93.8]; p=0.034 and median, 86.67; IQR, 76.9-100.0 vs. 77.78 [66.7-89.9]; p=0.028, respectively). No significant between-group differences were found in top-quality embryos (p=0.693) or the clinical pregnancy rate (65.9% vs. 61.8% in the consecutive vs. concomitant protocol, respectively). The median follicular fluid soluble Fas antigen level was significantly higher in the concomitant protocol (9,731.0 pg/mL; IQR, 6,004.5-10,807.6 vs. 6,350.2 pg/mL; IQR, 4,382.4-9,418.4; p=0.021). Conclusion: Personalized controlled ovarian stimulation using HP-hMG during the late follicular phase led to a significantly lower response, but did not affect the quality of ICSI.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.4
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• pp.352-361
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• 2021

Objective: The study assessed the developmental potential of germinal vesicle (GV) oocytes subjected to in vitro maturation (IVM) after prematuration culture with cilostamide (a phosphodiesterase-3 inhibitor) and the impact of cilostamide exposure on the morphology of meiosis II (MII) oocytes and subsequent embryo quality. Methods: In total, 994 oocytes were collected from 63 patients. Among 307 GV oocytes, 140 oocytes were selected for the experimental group and 130 oocytes for the control group. The denuded GV-stage oocytes were cultured for 6 hours with cilostamide in the experimental group and without cilostamide in the control group. After 6 hours, the oocytes in the experimental group were washed and transferred to fresh IVM medium. The maturational status of the oocytes in both groups was examined at 26, 36, and 48 hours. Fertilization was assessed at 18 hours post-intracytoplasmic sperm injection. Embryo quality was assessed on days 3 and 5. Results: In total, 92.1% of the oocytes remained in the GV stage, while 6.4% converted to the MI stage (p<0.01) after cilostamide exposure. In both groups, more MII oocytes were observed at 36 hours (25.8% vs. 21.5%) than at 26 hours (10.8% vs. 14.6%) and 48 hours (13% vs. 7.9%) (p>0.05). With the advent of cilostamide, blastocyst quality was better in the experimental group than in the control group (p<0.05). Conclusion: Cilostamide effectively blocked nuclear maturation and promoted cytoplasmic growth. Prematuration culture with cilostamide enabled synchronization between cytoplasmic and nuclear maturity, resulting in better blastocyst outcomes.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.293-297
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• 2016

For evaluating the boar semen quality, sperm motility is an important parameter because the movement of sperm indicates active metabolism, membrane integrity and fertilizing capacity. Phospholipase C zeta (PLCz) is important enzyme in spermatogenesis, but the effect has not been confirmed in pigs yet. Therefore, this study was aimed to analyze their association with sperm motility and kinematic characteristics. DNA samples from 124 Duroc pigs with records of sperm motility and kinematic characteristics [total motile spermatozoa (MOT), curvilinear velocity (VCL), straight-line velocity (VSL), the ratio between VSL and VCL (LIN), amplitude of lateral head displacement (ALH)] were subjected. A SNP in non-coding region of PLCz g.158 A > C was associated with MOT (p < 0.05), VCL (p < 0.01), LIN (p < 0.01) and ALH (p < 0.05) in Duroc population. Therefore, we suggest that the intron region of the porcine PLCz gene may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.2
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• pp.131-141
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• 2008

Objective: To compare the outcomes of intracytoplasmic sperm injection (ICSI) with fresh and cryopreserved-thawed testicular spermatozoa in patients with azoospermia. Methods: One hundred and nine cycles (66 couples) where ICSI was planned with fresh or cryopreserved-thawed testicular spermatozoa were included in this study; Ninety two cycles (61 couples) with fresh testicular spermatozoa (fresh group) and seventeen cycles (13 couples) with cryopreserved-thawed testicular spermatozoa (cryopreserved-thawed group). We compared ICSI outcomes such as fertilization rate, implantation rate, pregnancy rate and miscarriage rate, which were statistically analyzed using Mann-Whitney U test or Fisher's exact test, where appropriate. Results: In 9 out of the 92 cycles where ICSI was planned with fresh testicular spermatozoa, testicular spermatozoa could not be retrieved. Fertilization rate tended to be higher in the fresh group than in the cryopreserved-thawed group ($58.0{\pm}27.8%$ vs. $45.9{\pm}25.0%$, p=0.076). The number of high quality embryos was significantly higher in the fresh group ($0.9{\pm}1.2$ vs. $0.2{\pm}0.5$, p=0.002). However, there were no significant differences in clinical pregnancy rate, implantation rate and miscarriage rate between the two groups. Conclusion: The results of this study suggest that although the use of cryopreserved-thawed testicular sperm for ICSI in patients with azoospermia may reduce fertilization capacity and embryo quality, it may not affect pregnancy rate, implantation rate and miscarriage rate. If testicular sperm can be obtained before ICSI procedure, the use of cryopreserved-thawed testicular sperm may also avoid unnecessary controlled ovarian hyperstimulation and cancellation of oocyte retrieval when spermatozoa cannot be retrieved as well as damage on testicular function by repeated TESE.

• Journal of Embryo Transfer
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• v.33 no.1
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• pp.17-22
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• 2018

This study was conducted to determine the effect of pentoxifylline levels on sperm motility, survival rate, sperm membrane integrity of frozen semen and fresh-extended equine semen in Jeju cross-bred horses. As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw, the progressive motilities were $53.25{\pm}2.87$ (4mM pentoxifylline) and $50.28{\pm}2.14$ (8mM pentoxifylline) and significantly higher compared to the control group($40.09{\pm}5.15$) and other treatment group (16mM pentoxifylline, $41.27{\pm}2.82$). The progressive fast motility were $22.44{\pm}1.62$ (4mM pentoxifylline,) and $22.74{\pm}3.07$ (8mM pentoxifylline) and significantly higher compared to the control group ($13.47{\pm}1.48$) and other treatment group (16mM pentoxifylline, $14.66{\pm}3.68$) (p<0.05). As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw were $68.96{\pm}1.64$ (4mM pentoxifylline) and $67.90{\pm}6.72$ (8mM pentoxifylline) and significantly higher compared to the control group ($53.48{\pm}4.84$) and other treatment group (16mM pentoxifylline, $58.14{\pm}2.65$) (p<0.05). In conclusion, these results suggest that treatment groups with 4mM and 8mM pentoxifylline were higher compare to equine seperm mobility and the control group and treatment groups with more than 16mM pentoxifylline has a negative effect on sperm characteristics. After thawing, the total motility in post-thawed equine sperm has increased by 10 percent for 1 hour. these results suggest that pentoxifylline contributes to the improvement of the equine sperm motility and characteristics in post-thawed semen.