• Clinical and Experimental Reproductive Medicine
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• v.51 no.2
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• pp.120-124
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• 2024

Objective: Seasonal variations in semen quality are known to occur in temperate regions, but results regarding tropical areas remain inconclusive. The aim of this study was to determine whether monthly variations in semen parameters are present among men in a tropical region. Methods: Data were retrospectively collected from semen analyses of 3,000 men over a 10-year period, from 2012 to 2022. Analysis of variance and the independent-samples t-test were employed to observe variations in semen parameters throughout the entire period and between months, respectively. Results: The mean±standard deviation sperm concentration was significantly lower in June, at 42.5±31.4 million/mL, compared to other months. The highest sperm concentration was found in March, at 57.8±42.6 million/mL, constituting a mean difference of 15.3 million/mL between the lowest and highest concentrations. The total sperm count displayed a similar pattern of monthly variation, with a difference of 47.2 million between the highest and lowest months. No significant monthly differences were observed in other parameters, such as sperm motility, morphology, and semen volume. Conclusion: Significant monthly variations in sperm concentration and total sperm count were evident in this Sri Lankan population. March, which displayed the highest sperm counts, is in the spring in temperate regions, while the month with the lowest counts, July, is part of the summer. Fluctuations in photoperiod appear to most strongly influence these variations.

• Journal of Embryo Transfer
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• v.15 no.1
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• pp.85-94
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• 2000

These experiments were conducted to examine the effects of theophylline, pentoxifylline and heparin on frozen-thawed Hanwoo sperm for enhancing motility and viability of sperm. Frozen-thawed semen collected from one bull was treated in TALP(tyrode-albuminlactate-pyruvate) containing varous concentrations of theophylline and pentoxifylline. After incubated at 5% CO2 in air atmosphere for 6 hours, the motility of sperm after the treatments was characterized by CASA(computer aided semen analysis) system. When monitored notility(MOT) and curvilinear velocity(VCL), theophylline and pentoxifylline exerted their optimal action at the concentration of 30 mM and 3 mM, respectively. No difference of sperm motility was observed when the sperm was treated with both substances compared with a single treatment of each substance. Comparison was then made for evaluating the effect of theophylline and / or pentoxiophylline on the motility and viability of significant treatment effects of each substance, high MOT and VCL values were detected in sperm treated with theophylline. In the case of sperm viability examined by an eosin-nigrosin staining, however, a significant decrease was found after the combined treatment of theophylline+pentoxyphilline than after the treatment with heparin alone or no treatment(P<0.05). In conclusion, theophylline, pentoxiphylline or heparin can be used for enhancing the motional characteristics and viability of frozen thawed Hanwoo semen. Considering characteristics of these substances, theophyline may be useful in the artificial insemination system, which requires vigorous sperm motility. While, heparin supporting sperm viability in vitro can be effectively used for improving in vitro-fertilization system.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.1
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• pp.50-60
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• 2021

Objective: This study investigated the mRNA expression of gamma-aminobutyric acid (GABA) receptors in the sperm of oligoasthenoteratozoospermic (OAT) and teratozoospermic (TER) men compared to normozoospermic (NOR) men, as well as the relationships between GABA receptor expression and sperm parameters, fertilization rate, and embryo quality. Methods: The mRNA expression of GABA A-α1 and GABA B-R2 receptors in sperm was examined using reverse transcription-polymerase chain reaction in three groups of patients: NOR (n=32), OAT (n=22), and TER (n=45). The fertilization rate and embryo quality were assessed in 35 patients undergoing intracytoplasmic sperm injection (ICSI; 10 NOR, 10 OAT, and 15 TER men). Results: OAT men had significantly higher mRNA expression of GABA A-α1 and GABA B-R2 receptors in sperm than NOR men; however, the difference between TER and NOR men was not significant. High levels of these receptors were significantly correlated with low sperm concentration, motility, and morphology, as well as the rate of good-quality embryos (GQEs) at the cleavage stage after ICSI. Patients whose female partners had a >50% GQE rate at the cleavage stage had significantly lower levels of GABA A-α1 receptor expression than those whose partners had a ≤50% GQE rate. Conclusion: Our findings indicate that mRNA levels of GABA receptors in human sperm are correlated with poor sperm quality and associated with embryo development after ICSI treatment. The GABA A-α1 receptor in sperm has a stronger relationship with embryo quality at the cleavage stage than the GABA B-R2 receptor.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.2
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• pp.100-105
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• 2019

The purpose of this study was to evaluate the effect of addition of ethylene glycol, glycerol and sucrose to TCG (Tris, Citric Acid, Glucose, Egg Yolk) and DMSO Frozen. The extender containing Egg yolk concentration (10%, 20%) affects viability and acrosome morphology of rabbit sperm. Sperm viability was then assessed for the freezing extenders TCGD (Tris + Citricacid + Glucose + DMSO), TCGED (Tris + Citricacid + Glucose + Egg yolk + DMSO), TCGGD (Tris + Citricacid + Glucose + Glycerol + DMSO) and TCGSD Tris + Citricacid + Glucose + Sucrose + DMSO) during thawing at 38℃. for 20 seconds, respectively. TCG + 10% egg yolk (viability: 77.0 ± 0.8, NAI: 73.3 ± 0.9) was significantly (sperm viability and normal acrosome interaction (NAI)) higher than TCG + 20% egg yolk (70.7 ± 1.1, 70.0 ± 0.9) in the sperm normalcy analysis according to the yolk concentration. TCGGD (53.4 ± 0.1, 62.3 ± 0.4), TCGSD (61.3 ± 0.0, 67.1 ± 0.1) sperm viability and normal acrosome interaction (NAI) in frozen spermatozoa are TCGD (46.4 ± 2.8 and 56.3 ± 1. 4) and TCGED (23.0 ± 1.1 and 54.6 ± 1.4) extenders was thawed at 38℃ for 20 seconds. According to the results from each frozen bulking agent, sperm membrane integrity by hypotonic swelling test (HOST) analysis in TCGGD (59.8 ± 0.7), TCGSD (59.3 ± 0.5) was significantly high compared to other experimental groups (p < 0.05). In conclusion, these results suggested that TCGGD and TCGSD extenders enhance survivability of rabbit sperm after frozen-thawing.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.41-46
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• 2010

The objective of this study was to evaluated the efficiency of sperm cryosurvival using each extenders Triladyl and LEY containing egg yolk to the cryopreservation of separated sperm by percoll in miniature pig. The ejaculated semen from miniature pig was separated by 65% percoll and un-separated sperm as a control before freezing. The freezing of diluted semen added with Triladyl containing egg yolk and LEY solution (solution I: 11% Lactose or Triladyl + egg yolk; solution II: solution I + glycerol + OEP). Analysis of sperm ability was estimated by viability, capacitation acrosome reaction using chlortetracycline (CIC) the morphologic abnormality and hypoosmotic swelling test(HOST). The groups were designed that as separated sperm by Percoll with Triladyl(ST) or LEY(SL) for cryopreservation. And unseparated sperm with Triladyl(UT) or LEY(UL). As a results, the viability was higher significantly(p<0.05) in ST, SL, UT than UL extender. The morphologic abnormality was measured significantly (p<0.05) lower in ST than other extenders. The AR-patterned in CTC analysis was measured significantly(p<0.05) lower in SL and UL than other extenders. In conclusion, using Triladyl extender resulted in viability and morphology of separated sperm by percoll that were effective than using LEY extender, but it resulted in capacitation acrosome reaction was lower than using LEY extender.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.263-266
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• 2010

Prediction of semen's fertilizing ability used in artificial insemination (AI) is one of very important factors on pig reproductive performance. In vitro fertilization (IVF) has been used for indirect evaluation of sperm's fertilizing ability and it has been showed as highly correlated index. In swine industry, increasing interest in preservation of boar semen raises questions on the sperm motility from semen used in commercial AI centers. Mitochondria in sperm mid-piece generate the energy to support motility and could be an explanation of impaired fertility. Objective of this study was to suggest usable sperm motility to farms in measuring the effect of sperm motility and sperm abnormality on in vitro production of embryo in which sperm's fertilizing ability can be determined indirectly. Semen samples were provided from local AI center and used within 3 days after collection. Semen samples were divided by 4 different motile groups (>70%; 61~70%; 51~60%; <50%) using CASA (computer-assisted sperm analysis) on the days of IVF. Developmental rate to the blastocyst stage from over 61% motile sperm group showed significantly higher rate than below 60% motile sperm group ($16.5{\pm}0.7{\sim}18.4{\pm}0.8%$ vs $6.3{\pm}0.8{\sim}11.5{\pm}0.7%$, p<0.05). In experiment to determine the relationship between sperm motility and viability and abnormality, over 61% motile sperm groups showed significantly higher viability rate compared to below 60% motile sperm groups ($84.8{\pm}4.0{\sim}88.1{\pm}4.0%$ vs $69.1{\pm}4.0{\sim}74.2{\pm}4.0%$, p<0.05). On the other hand, morphological sperm abnormality showed significantly higher in over 70% motile sperm group ($10.2{\pm}2.2$ vs $16.0{\pm}2.2{\sim}21.0{\pm}2.2%$, p<0.05). In experiment to find the correlation between sperm motility of 4 different motile groups and amount of mitochondria, lower motility group also showed lower level of mitochondria (p<0.05). The mitochondria parameter used in this study showed another possibility to differentiate the sperm motility. Taken together, because below 60% motile semen used in AI reduce the fertility, AI centers should provide the over 60% motile sperm to the farms at the time of AI.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.265-271
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• 2013

Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG $0.2{\mu}M$, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.10
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• pp.1417-1425
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• 2014

The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not sufficient to select normal and fertilizable spermatozoa. In conclusion, the present study suggests that the acrosome staining and detailed motion characteristics such as progressiveness, VCL, and VSL should be included in determining semen quality together with primary parameters for successful AI and high breeding performance in the swine industry.

• Development and Reproduction
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• v.21 no.3
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• pp.229-235
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• 2017

In the field of reproductive medicine, assessment of sperm motility is a key factor for achieving successful artificial insemination, in vitro fertilization, or intracellular sperm injection. In this study, the motility of boar sperms was estimated using real-time imaging via confocal microscopy. To confirm this confocal imaging method, flagellar beats and whiplash-like movement angles were compared between fresh and low-temperature-preserved ($17^{\circ}C$ for 24 h) porcine sperms. Low-temperature preservation reduced the number of flagellar beats from $11.0{\pm}2.3beats/s$ (fresh sperm) to $5.7{\pm}1.8beats/s$ and increased the flagellar bending angle from $19.8^{\circ}{\pm}13.8^{\circ}$ (fresh) to $30.6^{\circ}{\pm}15.6^{\circ}$. These data suggest that sperm activity can be assessed using confocal microscopy. The observed motility patterns could be used to develop a sperm evaluation index and automated confocal microscopic sperm motility analysis techniques.

• Environmental Analysis Health and Toxicology
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• v.19 no.2
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• pp.177-182
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• 2004

Alcohol consumption and exposure to endocrine disruptors and industrial solvents have been implicated in impaired spermatogenesis, increase in the incidence of malformed sperm and decrease in the percentage of moving sperm. The aim of this study was to determine and compare the direct effects of some organic solvents(bisphenol A; BPA, dibutyl phthalate; DBP, formaldehyde; HCHO, dimethylsulphoxide; DMSO, ethanol) on mucus penetration distance, motility and survival rate of human sperm in vitro. Semen samples from 3 health subjects were prepared using swim-up method and 0.0005-0.5% organic solvents were added to the test medium. BPA, DBP, HCHO and DMSO produced significant decreases in the motility and survival rate with a different potency. The most potent inhibition of mucus penetration distance, motility and survival rate was observed after exposure to HCHO. A concentration of 0.0005% HCHO significantly inhibited sperm motility. When ethanol m.: added directly to sperm, at concentrations equivalent to that in serum after heavy drinking, these damaging effects were lowest compared with other solvents. Present study shows that each compound has different toxic potency to human sperm and we need special caution for the use of HCHO.