• Reproductive and Developmental Biology
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• 제40권1호
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• pp.1-5
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• 2016

This study investigated the effects of L-carnitine (LC) and nicotinic acid (NA) on sperm viability during liquid storage at $18^{\circ}C$ in miniature pigs. $10{\mu}M$ LC and 30 mM NA, combined LC and NA (LN) were treated in fresh semen for 3, 7, and 10 days. In results, sperm survival increased in NA- and LN-treated semen on 7 and 10 days (p<0.05), mitochondrial integrity of live sperm increased in LN-treated semen on 7 days (p<0.05), but not NA-treated semen. In addition, we examined the acrosome reaction of sperm in miniature pigs. LC and NA did not influence on acrosome reaction of boar sperm. In conclusion, LC and NA effectively maintained the viability and quality of sperm during long-term storage in miniature pigs, suggesting that the combined LN may be useful for improving the semen extender for long-term liquid storage in pigs.

• 생명과학회지
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• 제29권4호
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• pp.395-401
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• 2019

Phthalate는 내분비 교란물질로 호르몬의 변화, 에스토젠, 앤드로젠, 갑상선 호르몬의 분비를 방해한다. 또한, 인간과 동물에서 심혈관질환, 대사작용, 면역 및 번식체계에 영향을 끼친다. Curcumin은 항산화물질로 항염증 활성 및 항암작용에 영향을 미치는 것으로 알려져 있다. 본 연구는 phthalate가 정자의 운동성, 생존율, 미토콘드리아 활성 및 세포막 기능에 미치는 영향을 알아보고자 실시하였다. 또한, curcumin을 처리하여 phthalate에 노출된 정자에 미치는 영향을 분석하였다. 정자의 운동성과 생존율은 di-n-butyl phthalate (DBP), mono-n-butyl phthalate (MBP) 및 di-2-ethylhexyl phthalate (DEHP)을 처리하였을 때 감소하였다(p<0.05). Phthalate는 정자의 미토콘드리아 활성 및 세포막의 기능을 감소시켰다(p<0.05). 그러나, 정자의 운동성과 생존율은 curcumin을 처리하지 않은 것보다 처리한 정자에서 높게 나타났으며(p<0.05), 정자의 미토콘드리아 활성 및 세포막 기능에서도 높게 나타났다(p<0.05). 결론적으로, phthalate는 정자의 생존율과 세포의 기능에 영향을 미칠 수 있고, 이로부터 세포의 기능을 보호하기 위해서는 curcumin의 처리가 필요 할 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권7호
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• pp.924-929
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• 2004

The objective of this study were to investigate the effect of ejaculation frequency on semen characteristics and to establish a method for quick assessment of sperm concentration in TCC using packed cell volume (PCV) as the parameter (Trial 1). Eighty senior roosters, averaging 61 wk-old, were used and the sperm concentrations were determined using a hemacytometer. The PCV value was measured in a capillary (0.75 mm in inner diameter) by centrifugation. A simple linear regression analysis suggested that the sperm concentrations were significantly correlated with PCV values (r=0.62, p<0.001). Trial 2 was conducted to determine the optimal ejaculation frequency of TCC roosters in a weekly semen collection program. The male birds were subjected to 1, 2, 3 or 6 ejaculations per week for four consecutive weeks and semen characteristics including ejaculation volume (EV, mL), sperm motility (%), PCV (%), sperm concentration (ESC, $\times$10$^{9}$/mL), weekly sperm production (WSP, $\times$10$^{9}$/wk) and average motile sperm numbers (AMSN, $\times$10$^{9}$/ejac) were determined. Average EV was greater in the group with 3 ejac/wk than with only 1 ejac/wk in weeks 1 and 3 of the collection period. WSP increased with ejaculation frequency during the first 3 weeks of collection (p<0.05). Sperm motility was better in the birds with 6 ejac/wk than in single ejaculation group for the first 2 wk and no significant differences were found for the last 2 wk of study. In contrast, the PCV value showed a trend of reduction for the first 2 wks in the 6 ejac/wk group. Surprisingly, no significant differences were detected in the AMSN among treatment groups. The weekly motile sperm production (WMSP) increased with ejaculation frequency. Based on our observation, PCV values could be used for a quick estimation of sperm concentration and an intensive semen collection program enhanced weekly sperm production in TCC roosters.

• 한국발생생물학회지:발생과생식
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• 제19권1호
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• pp.1-10
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• 2015

The purpose of the present study was to examine the seminiferous epithelium cycle of Bombina orientalis using a light microscope. The cycle was divided into a total of 10 stages, according to the morphological characteristics of the cells. The spermatogenetic cells included primary spermatogonia, secondary spermatogonia, primary spermatocytes, secondary spermatocytes, spermatid and sperm. At stage I, the primary spermatogonia was located closer to basal lamina of the seminiferous tubule without spermatocyst formations. Especially at the stage II, the secondary spermatogonia were located in the spermatocyst. The primary and secondary spermatocytes were found from stages III to VI. The secondary spermatocytes were smaller in size than the primary spermatocytes, but they had thicker nucleoplasm and smaller nuclei. The round-shaped, early sperm cells were formed in stage VII, and further divided at stage VIII to have more concentrated nucleoplasm before division to matured sperm cells. At stage X, the matured sperm cells emerged from the spermatocyst. Considering the above results, this study presented the special characteristics in the generation and type of sperm formation. The germ cell formation occurred in various stages, like the perspectives of Franca et al (1999), ultimately, providing taxonomically useful information.

• Asian-Australasian Journal of Animal Sciences
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• 제24권2호
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• pp.181-189
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• 2011

Cryopreservation protocols induce partially irreversible damage to mammalian sperm plasma membranes. Previous studies have indicated that adding cholesterol to the plasma membrane, as cholesterol-loaded-cyclodextrins, improves cryosurvival of sperm. Therefore, the purpose of this study was to determine if treating sperm of Markhoz bucks with cholesterol-loaded-cyclodextrins (CLC) (0, 0.75, 1.5, 2.25 and 3 mg/ml diluted $240{\times}10^6$ sperm/ml) in Tris-citric acid-glucose diluents with and without egg yolk (containing 5% glycerol) would improve the post-thaw sperm quality. The motion characteristics were evaluated with a Computer Assisted System Analyzer (CASA); acrosome integrity and vitality were measured with the triple-stain technique. Samples were recovered before and after freezing by means of putting straws into $37^{\circ}C$ water for 30 sec and then parameters were assessed. The results showed that the treatments significantly affected motility, progressive motility, recovery rate, curvilinear velocity, beat cross frequency, live sperm with reacted acrosome, live sperm with unreacted acrosome, dead sperm with reacted acrosorne, and dead sperm with unreacted acrosome during freezing (p<0.05). However; no significant differences were found for average path velocity, straight line velocity, amplitude of lateral head displacement, straightness and linearity (p>0.05). The best results were observed for extender containing 2.25 mg/ml ($240{\times}10^6$ sperm/ml) CLC supplemented with 2.6% egg yolk. In conclusion, the findings of this study indicate improved Markhoz sperm viability and motility following treatment in the presence of egg yolk.

• Asian-Australasian Journal of Animal Sciences
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• 제33권11호
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• pp.1763-1769
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• 2020

Objective: This study was conducted to investigate the effect of L-carnitine on the pig semen characteristics during storage. Methods: Spermatozoa samples were examined for spermatozoa quality and then randomly divided into 5 groups: 0 (control), 12.5, 25, 50, and 100 mM L-carnitine. Sperm motility, plasma membrane integrity and antioxidant parameters (total reactive oxygen species, total antioxidant capacity, and malondialdehyde) were evaluated after 0, 3, 5, and 10 day cooled-storage at 17℃. Moreover, ATP content, mitochondria activity as well as sperm-binding and in vitro fertilizing ability of preserved boar sperm were also investigated. Results: Supplementation with 50 mM L-carnitine could effectively maintain boar sperm quality parameters such as sperm motility and membrane integrity. Besides, we found that L-carnitine had positive effects on boar sperm quality mainly through improving antioxidant capacities and enhancing ATP content and mitochondria activity. Interestingly, by assessing the effect of L-carnitine on sperm fertility and developmental potential, we discovered that the extender containing L-carnitine could improve sperm quality and increase the number of sperms bounding to zona pellucida, without improving in vitro fertility and development potential. Conclusion: These findings suggested that the proper addition of L-carnitine to the semen extender improved boar sperm quality during liquid storage at 17℃.

• 한국동물생명공학회지
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• 제37권2호
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• pp.130-135
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• 2022

Epididymal sperm cryopreservation provides a potential method for preserving genetic material from males of endangered species. This pilot study was conducted to develop a freezing method for tiger epididymal sperm. We evaluated post-thaw sperm condition using testes with intact epididymides obtained from a Siberian tiger (Panthera tigris altaica) after castration. The epididymis was chopped in Tyrode's albumin-lactate-pyruvate 1x and incubated at 5% CO₂, 95% air for 10 min. The Percoll separation density gradient method was used for selective recovery of motile spermatozoa after sperm collection using a cell strainer. The spermatozoa were diluted with modified Norwegian extender supplemented with 20 mM trehalose (extender 1) and subsequent extender 2 (extender 1 with 10% glycerol) and frozen using LN₂ vapor. After thawing at 37℃ for 25 s, Isolate^® solution was used for more effective recovery of live sperm. Sperm motility (computerized assisted sperm analysis, CASA), viability (SYBR-14 and Propidium Iodide) and acrosome integrity (Pisum sativum agglutinin with FITC) were evaluated. The motility of tiger epididymal spermatozoa was 40.1 ± 2.0%, and progressively motile sperm comprised 32.7 ± 2.3%. Viability was 56.3 ± 1.6% and acrosome integrity was 62.3 ± 4.4%. Cryopreservation of tiger epididymal sperm using a modified Norwegian extender and density gradient method could be effective to obtain functional spermatozoa for future assisted reproductive practices in endangered species.

• 한국가축번식학회지
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• 제21권4호
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• pp.405-409
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• 1997

This study was carried out to investigate the general characteristics such as semen volume, pH, sperm motility and sperm concentration of the semen collected from Korean Jindo dogs by the mothod of Digital manipulation of penis, and the effect of temperature and preservation time on motility of fresh semen. Multiple ejaculates were collected from four male Korean Jindo dogs. The results obtained in this experiment were as follows : 1. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the second fraction and the small volume of third fraction from the ejaculate were 3.29ml, 6.30, 96.70% and 1.64$\times$108 cells/ml, respectively. 2. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculate were 1.16ml, 6.10, 6.67% and 5.07$\times$105cells/ml. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the second fraction from the ejaculate were 2.30ml, 6.33, 97.66% and 1.92$\times$108cells/ml. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the third fraction from the ejaculate were 3.24ml, 6.51, 93.33% and 3.13$\times$107cells/ml. 3. Motility of fresh semen during preservation were higher at 17$^{\circ}C$ than at 5$^{\circ}C$ or 36$^{\circ}C$. When preservation temeprature was 17$^{\circ}C$, motility were 95.75% at 1 h, 90.00% at 6 h, 84.25% at 12 h, 68.00% at 18 h, 36.25% at 24 h and 28.75% at 30 h, respectively.

• 농업과학연구
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• 제14권2호
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• pp.413-421
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• 1987

토끼에서 bovine serum albumin gradients로 성(性)의 전선택(前選擇)이 가능(可能)한지를 알아보기 위하여 6%, 10% 및 20%의 농도(濃度)를 달리한 bovine serum albumin을 사용한 방법(方法)으로 고활력정자(高活力精子)를 분리수집하여 정자(精子)의 성상(性狀)과 분리된 정액(精液)의 인공수정시(人工授精時)에 있어서 자토(仔兎)의 성비(性比)를 비교관찰한 결과는 다음과 같다. 1. Bovine serum albumin을 사용하여 분리(分離)된 정자(精子)는 임신율(妊娠率)에 있어서는 대조군에 비해 높은 성적(成績)을 나타내었으나, 자토(仔兎)의 성비(性比)에 있어서는 변화를 일으키지 못하였다. 2. Bovine serum albumin에 의해 분리(分離)된 정자(精子)는 대조군에 비하여 운동성(運動性), 정상정자율(正常精子率) 및 전진운동성(前進運動性)이 현저히 높았으며, 정자회수율(精子回數率)은 6%일 때 가장 높았다. 3. 원정액(原精液)으로부터 bovine serum albumin을 사용하여 고활력정자(高活力精子)를 분리(分離)한 후(後) 냉동(冷凍)한 정액(精液)의 융해후(融解後) 정자운동성(精子運動性), 정상정자율(正常精子率) 및 전진운동성(前進運動性)은 대조군에 비하여 현저히 높았다.

• Clinical and Experimental Reproductive Medicine
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• 제44권3호
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• pp.132-140
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• 2017

Objective: Correlations between semen parameters and sperm DNA fragmentation index (DFI) were investigated to identify characteristics of sperm without DNA damage that could be used in selecting sperm for intracytoplasmic sperm injection (ICSI). Pregnancy outcomes were compared to determine whether in vitro fertilization (IVF) or ICSI is a better choice for patients who have sperm with a high-DFI. Methods: Semen analysis was carried out in 388 patients who visited our IVF center for the first time to investigate correlations between sperm DFI and semen parameters. In addition, 1,102 IVF cycles in 867 patients were carried out in the present study; 921 cycles in the low-DFI group (DFI < 30%) and 181 cycles in the high-DFI group ($DFI{\geq}30%$). Both the low- and high-DFI groups were subdivided into IVF and ICSI cycle groups. Results: Sperm DFI showed significant inverse correlations with sperm motility (r = -0.435, p< 0.001) and morphology (r = -0.153, p< 0.05). Sperm DFI also showed significant correlations with rapid motility (r = -0.436, p< 0.001), and the kinetic parameters of average-path velocity (r = -0.403) and linearity (r = -0.412). Although there was no significant difference in the pregnancy rates between IVF (48.6%) and ICSI (44.8%) in the low-DFI group, the pregnancy rate of ICSI cycles (44.8%, p< 0.05) was significantly higher than IVF cycles (25.0%) in the high-DFI group. No significant difference was observed in the abortion rates between the low-DFI (52 of 921, 5.6%) and high-DFI groups (7 of 181, 3.8%). Conclusion: ICSI is a better choice than IVF for improving the pregnancy outcomes of patients who have sperm with a high DFI.