• 대한생식의학회:학술대회논문집
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• 대한불임학회 2000년도 제39차 춘계 학술대회
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• pp.37-42
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• 2000

Objective: To determine whether particular semen characteristics in various clinical diagnoses of infertility are associated with high oxidative stress and whether any group of infertile men is more likely to have high seminal oxidative stress. Reactive oxygen species (ROS) play an important role in sperm physiological functions, but elevated levels of ROS or oxidative stress are related to male infertility. Design: Measurement of sperm concentration, motility, morphology, seminal ROS, and total antioxidant capacity (TAC) in patients seeking infertility treatment and controls. Setting: Male infertility clinic of a tertiary care center. Patient(s): One hundred sixty-seven infertile patients and 19 controls. Intervention(s): None. Main Outcome Measure(s): Semen characteristics, seminal ROS, and TAC in samples from patients with various clinical diagnoses and controls. Result(s): Fifteen patients (9.0%) were Endtz positive and 152(91.0%) Endtz negative. Sperm concentration, motility, and morphology were significantly reduced in all groups compared with the controls (P = .02), except in varicocele associated With infection group. Mean (${\pm}$SD) ROS levels in patient groups ranged from 2.2 ${\pm}$ 0.13 to 3.2 ${\pm}$ 0.35, signilicantly higher than controls (1.3 ${\pm}$ 0.3; P<.005). Patient groups had a significantly lower mean (${\pm}$SD) TAC from 1014.75 ${\pm}$ 79.22 to 1173.05 ${\pm}$ 58.07 than controls (1653 ${\pm}$ 115.28, P<.001), except ill the vaseclony reversal group (1532.02 ${\pm}$ 74.24). Sperm concentration was negatively correlated with ROS both overall and within all groups (P${\leq}$.007), with the exception of idiopathic infertility. Conclusion(s): Irrespective of the clinical diagnosis and semen characteristics, the presence of seminal oxidative stress in infertile men suggests its role in the pathophysiology of infertility. Medical or surgical treatments for infertility in these men should include strategies to reduce oxidative stress.

• 한국수정란이식학회지
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• 제21권1호
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• pp.69-75
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• 2006

본 연구는 제주도 축산진흥원과 농촌진흥청 난지농업연구소에서 사육되고 있는 제주마 5두를 사용하여 인공질을 이용, 정액을 채취하여 정액의 일반성상 및 CASA를 이용하여 정자의 운동성과 생사 염색으로 기형률을 조사하였다. 제주마 정액의 일반 특성으로 총 정액량은 평균 42.5ml이었으며 평균 pH는 7.3, 평균 정자농도는 평균 $198.5X10^6/m1$ 이었다. 정자의 운동성은 $64.3{\pm}23.2%$이며, motility parameters 별로 VAP $70.4{\pm}28.7{\mu}m/s,\;VSL\;69.6{\pm}28.9{\mu}m/s,\;VCL\;94.1{\pm}30.0{\mu}m/s,\;ALH\;2.3{\pm}0.7{\mu}m/s,\;BCF\;7.6{\pm}1.1Hz,\;STR\;99.1{\pm}1.2%,\;LIN\;77.1{\pm}12.7%$로 나타났다. 정자 부위별 기형률은 두부가 평균 4 %, 경부가 평균 20 %, 미부가 평균 4 %로 나타났다. 이 연구를 통해 천연기념물로 지정되어 있는 제주마의 유전자원 보존과 증식을 위한 인공수정기술의 실용화를 위한 기초 자료를 제공하고자 한다.

• 한국수정란이식학회지
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• 제28권4호
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• pp.349-353
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• 2013

The objective of this study was to compare the effect of semen extenders on the sperm motility, viability, acrosome integrity and functional integrity of plasma membrane (HOST: hypo-osmotic swelling test) during liquid preservation of Korean Native boar semen. In this experiment, semen was diluted in Androhep plus, Beltsville Thawing Solution (BTS), ModenaTM, Seminark and Vitasem LD. Sperm-rich fractions were collected from three Korean Native boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different semen extenders. Semen samples were stored at $17^{\circ}C$ for 96 hours. On everyday (0, 24, 48, 72, 96 h) after storage, the sperm characteristics relevant for fertility, such as sperm motility, viability, acrosome integrity and HOST positive were evaluated. The motility of spermatozoa stored in different extenders was no significantly different among other extenders (P>0.05). Also, no difference was observed among samples processed with different extenders in the percentage of sperm viability, acrosome integrity and HOST positive. All extenders maintained a high percentage (70%) of sperm motility, viability and acrosome integrity through 96 h of storage. The result of this study show that there was no significant differences among extenders in their capacity to preserve motility, viability and membrane integrity of spermatozoa from normal, fertile Korean Native boars for 96 h of liquid preservation at $17^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• 제16권2호
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• pp.165-171
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• 2003

Sixty semen ejaculates collected at weekly interval from four Murrah Buffalo bulls over a period of seven months (Nov.1999 to May 2000) were used in the present study. Three buffer medium (sodium citrate, TES and Tris) were used for soaking of sephadex. Three grades of sephadex (G-15,G-100, and G-200) were used for preparation of columns. Columns of three different height (one, two and three cm) were used for separation of semen. Twenty semen ejaculates were used in each project. In the first experiment each semen ejaculates was divided into four parts. One part was kept as control and other three parts were passed thought one cm column of sephadex G-15 prepared in three different buffers. There was significant (p<0.05) increase in percent progressive sperm motility and percent live spermatozoa and decrease in percent abnormal spermatozoa and percent spermatozoa with damaged acrosome as well as sperm numbers after filtration through all the three columns. Sperm quality obtained in the filtrate of column prepared in Tris buffer was better in comparison to other two buffers. So the Tris buffer was used in the second trial. Twenty semen ejaculates were used in this experiment. Each semen ejaculate was divided into four parts. One part was kept as control (non-filtered) and other three parts were passed through columns of different grade of sephadex (G-15, G-100 and G-200). Progressive sperm motility and live sperm percentage improved significantly while decline in percent abnormal spermatozoa and percent spermatozoa with damaged acrosome and sperm concentration was observed after filtration through all the columns as compared to control (non-filtered) semen. Since post filtration quality of semen was better in the sephadex G-100 column, therefore it was selected for the next experiment. In third experiment, Tris buffer and sephadex G-100 were used for preparing columns of different height (one, two and three cm) and twenty semen ejaculates were filtered. The quality characteristics of semen (percent progressive sperm motility, percent live spermatozoa and sperm concentration) after filtration through one cm column were significantly (p<0.05) higher than after filtration through columns of two and three cm height. However non -significant (p>0.05) difference due to height of columns was observed for percent abnormal and percent damaged acrosome but 1 cm column comparatively gave better result than of 2 and 3 cm column height.

• 한국수정란이식학회지
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• 제28권2호
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• pp.121-125
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• 2013

This study was carried out to investigate the general characteristics of semen such as semen volume, pH, sperm motility and sperm concentration of the semen collected from Shih Tzu dogs (age of 24 to 48 months, weight of 4 to 8 kg) by using the method of digital manipulation of the penis. The effect of preservation temperature and time on motility of fresh semen was also investigated in the present study. Semen was collected for 16 times from 4 male Shih Tzu dogs by multiple ejaculations (four times ejaculation per dog). The average of semen volume, semen pH, sperm motility and sperm concentration of the second fraction containing small volume of the initial third fraction per ejaculation were $2.11{\pm}0.31$ ml, $6.25{\pm}0.07$, $97.59{\pm}1.03%$ and $2.05{\pm}0.14{\times}10^8$ cells/ml, respectively. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculation were $1.12{\pm}0.15$ ml, $5.99{\pm}0.14$, $16.09{\pm}6.18%$ and $5.16{\pm}2.03{\times}10^5$ cells/ml, respectively. Those of second fraction were $2.07{\pm}0.29$ ml, $6.36{\pm}0.13$, $97.31{\pm}1.36%$ and $2.15{\pm}0.30{\times}10^8$ cells/ml, respectively. Those of third fraction were $2.60{\pm}0.29$ ml, $6.63{\pm}0.08$, $95.72{\pm}1.61%$ and $6.03{\pm}1.83{\times}10^7$ cells/ml, respectively. Sperm motility was significantly higher at $17^{\circ}C$ preservation temperature than at $5^{\circ}C$ or $36^{\circ}C$ during preservation period except 1 h preservation (P<0.05). When preservation temperature was $17^{\circ}C$, sperm motility was $96.69{\pm}1.49%$ at 1 h, $91.38{\pm}1.90%$ at 6 h, $88.38{\pm}2.34%$ at 12 h, $78.13{\pm}4.58%$ at 18 h, $58.44{\pm}8.57%$ at 24 h and $29.56{\pm}5.06%$ at 30 h, respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제23권10호
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• pp.1276-1281
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• 2010

The present study was conducted to investigate the effects of butylated hydroxytoluene (BHT) supplementation on diluted, cooled and frozen-thawed ram spermatozoa. After primary evaluation of collected ejaculates, only semen samples with motility of more than 70% and sperm concentration higher than $3{\times}10^3$ sperm/ml were used for cryopreservation. The selected semen samples were then pooled and diluted 1:4 with Tris Citrate Fructose Yolk (TCFY) extender supplemented with different concentrations of BHT (0.5, 10, 2.0 and 3.0 mM). As the control, semen was diluted and frozen in the diluent without BHT. Motility, progressive motility, viability, membranes and acrosome integrity were evaluated after dilution (part 1), cooling (part 2) and freezing and thawing (part 3). The results of the first part of the experiment showed that there were no significant difference between treatments in the motility, progressive motility, viability, membranes and acrosome integrity of spermatozoa, but the results with 2.0 mM BHT were slightly better than obtained with other levels of BHT and control extender. Significantly better results (p<0.05) were observed in the second part of the experiment for cooled spermatozoa characteristics, when extender was supplemented with 2.0 and 3.0 mM BHT. Furthermore, the results obtained in the third part of the experiment indicated that, after freezing and thawing, all evaluated semen characteristics were improved significantly (p<0.05) by increasing BHT levels, with the best results obtained for extender containing 2 mM BHT. Comparison of these results with those of control diluent, the effects of supplementation were significantly (p<0.01) better. However, the higher concentration of BHT (3.0 mM) reduced the motility, acrosomal integrity, viability and hypo-osmotic swelling response of spermatozoa compared to extender containing 2.0 mM BHT. In conclusion, the results obtained in this study showed that the semen quality of rams was improved when BHT was added to extender used before the freezing process.

• Asian-Australasian Journal of Animal Sciences
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• 제33권9호
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• pp.1411-1420
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• 2020

Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand. Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station; Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLC-selection had a favorable effect on PRO, VAP, and WOB that differed among seasons. Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.27-32
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• 2012

본 연구는 강원도 영동지방의 7개 시 군(강릉, 동해, 태백, 삼척, 속초, 양양, 고성)의 93개 농가에서 사육 중인 1,655두의 암소의 인공수정에 사용된 한우보증씨수소(KPN, Korean proven bulls number) 정액을 대상으로 인공수정 실패율과 동결정액의 성상 분석에 대해 실시하였다. 암소 인공수정에 사용된 KPN 정액의 인공수정 실패율 범위는 27.6~77.2%로 나타나, 씨수소 간의 인공수정 실패율 차이가 52.9%로 크게 나타났으며, 이중 KPN506이 27.6%로 가장 낮은 인공수정 실패율을, KPN593이 77.2%의 가장 높은 인공수정 실패율을 나타냈다. 또한, KPN과 암소의 인공수정 실패율은 0.2941의 정의 상관관계를 보여 암소의 인공수정에 있어 KPN 정액의 실패율은 상당한 영향을 미치는 것으로 사료된다. 유세포 분석기(flow cytometry)를 이용한 KPN593과 KPN637의 동결정액 성상분석결과에서 인공수정 실패율이 높은 KPN593의 생존율(viability rate)이 30.49%로 KPN637(37.34%)에 비해 유의적으로 낮은 결과를 보였으며($p$<0.05), 유의적인 차이를 보이진 않았지만 첨체 이상율 평가(ruptured acrosome rate)에서도 KPN593 (28.38%)이 KPN637(21.37%) 보다 높은 수치를 나타내었다. 이 결과로 보아 KPN 정액의 번식능력 및 KPN 동결정액의 품질은 암소 인공수정 실패와 매우 밀접한 관련이 있으며, 암소의 수태율 증가를 위해 인공수정 실패율이 높은 KPN은 피해야 하며, 향후 농가 이익을 위해 한우 보증씨 수소의 번식능력과 동결정액의 정확한 성상평가가 이루어져야 할 것으로 사료된다.

• 한국가축번식학회지
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• 제26권3호
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• pp.253-261
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• 2002

본 연구는 진도개 동결정액 제조기술을 정립하기 위하여 진도개 정액성상과 동결 전후 정액의 활력과 생존율 및 CASAs를 이용한 운동성 등에 대하여 조사하였고, 백구와 황구간, 개체간의 정액성상과 내동성을 비교, 조사하였다. 이상의 연구결과는 다음과 같다. 1. 총 63회 정액채취 후 신선정액의 평균 정액량 3.8 $m\ell$, 농도 145.6$\times$$10^{6}$$m\ell$, 총정자수 396.2$\times$08/$m\ell$, 전진운동율 79.7% 및 생존율 89.5% 였다. 황구와 백구간에는 황구가, 개체간에는 황구 2호가 정자농도, 총정자수, 전진운동정자율 및 생존율 등의 정액성상에서 유의적으로 우수하였다(P<0.05). 2. 동결전.후 정자의 전진운동율과 생존율을 46회 조사한 결과 동결전 73.5%와 82.3%를, 동결후 51.1%와 64.9%를 나타내 동결과정이 전진운동율과 생존율에 영향이 있었으며, 역시 황구와 백구간에는 황구가, 개체간에는 황구 2호가 동결전후 전진운동율과 생존율에서 유의적인 차이가 인정되었다 (P<0.05). 3. 동결.융해정자의 보다 객관적인 평가를 위하여 CASA system을 이용한 총 44회 평가한 결과, 생존율 65.6%, 전진운동율 54.8%, VAP 75.3 $\mu\textrm{m}$/sec, VCL 90.0 $\mu\textrm{m}$/sec, VSL 69.4 $\mu\textrm{m}$/sec 및 ALH 4.4 $\mu\textrm{m}$로 동결 융해 정액의 운동성은 양호하였고, 황구와 백구간의 운동성에는 유의적인 차이가 없었으나, 개체간에는 역시 황구 2호가 운동성이 우수하여 유의적인 차이가 인정되었다 (P<0.05). 4. 채취된 정액중 백구는 46%(13/28), 황구는 94%(33/35)가 동결정액 제조가 가능해 황구가 내동성이 좋았으며, 전체적으로 73%(46/63) 동결정액 제조가 가능하였다. 결론적으로 진도개의 동결정액을 제조하기 위하여 정액성상 및 동결 전후 운동성을 조사한 결과 동결정액 제조와 생산체계의 구축이 가능하였으며, 황구와 백구간, 개체간의 정액성상과 동결전후의 운동성에 차이가 인정되므로 정액성상과 내동성을 고려한 종견선발 체계가 필요하다고 사려되었다.

• 한국수정란이식학회지
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• 제29권4호
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• pp.385-391
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• 2014

Objective of this study was to investigate the effect of nicotinic acid (NA) on the characteristics in fresh semen of miniature pig. We evaluated viability, acrosome reaction and mitochondrial integrity of sperm on 0, 3, 7 and 10 days during storage period with nicotinic acid. As results, the survival rate of sperm in 15 mM NA (day 3, $87.8{\pm}1.2%$; day 5, $84.0{\pm}2.7%$; day 7, $82.2{\pm}0.9%$) and 30 mM NA (day 3, $87.7{\pm}0.3%$; day 5, $84.4{\pm}2.5%$; day 7, $82.3{\pm}0.7%$) groups were higher than control and 5 mM NA groups in 3, 7 and 10 days of semen storage. The NA-treated sperm on 10 day was used day for observing acrosome integrity. The survival sperm with acrosome reaction was higher in 30 mM NA group (day 3, $2.7{\pm}0.2%$; day 5, $3.3{\pm}0.6%$; day 7, $11.4{\pm}0.3%$) than in the control, significantly (P<0.05). Moreover, the live sperm with mitochondrial integrity was higher in whole treatment groups of NA than control group, significantly (P<0.05). Specially, most mitochondrial integrity on 10 day of semen storage was significantly higher in 30 mM NA group ($90.2{\pm}1.6%$) than other treatment groups (control, $81.8{\pm}3.1%$; 5 mM NA, $83.4{\pm}3.0%$; 15 mM NA, $89.1{\pm}0.7%$, P<0.05). In conclusion, supplement of NA in liquid semen of miniature pig can improve and maintain semen quality, such as viability, acrosome reaction, and mitochondria integrity.