• Journal of Life Science
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• v.29 no.4
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• pp.395-401
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• 2019

Phthalate is a chemical endocrine disrupter and interfere with the action of hormones, estrogens, androgens and thyroid hormones. It also affect cardiovascular, metabolic, immune and reproductive system in the human and animals. Curcumin is antioxidant, anti-inflammatory activity and -cancer properties in the human. We studied whether phthalates damage viability, mitochondrial activity and membrane integrity of sperm in boar semen. We also treated curcumin with/without phthalates in the boar semen. Fresh boar semen was treated with phthalates and/or curcumin for examining sperm characteristics. Sperm characteristics, sperm motility, viability, mitochondrial activity, and membrane integrity were determined during storage of boar semen. Sperm motility and viability in dose-dependent manner decreased by di-n-butyl phthalate (DBP), mono-n-butyl phthalate (MBP) and di-2-ethylhexyl phthalate (DEHP, p<0.05). Phthalates also decreased mitochondrial activity and membrane integrity of sperm (p<0.05). However, sperm motility and viability were higher than untreated-curcumin when DBP, MBP and DEHP treated with a curcumin in boar semen (p<0.05). Mitochondrial activity and membrane integrity of sperm were higher in DBP- and MBP-treated semen with curcumin (p<0.05). In conclusion, phthalates can damage sperm viability and quality during the boar semen storage, and curcumin may protect the boar sperms from phthalates during storage term.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.7
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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.

• Development and Reproduction
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• v.19 no.1
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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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• v.24 no.2
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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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• v.33 no.11
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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℃.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.2
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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.

• Korean Journal of Animal Reproduction
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• v.21 no.4
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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.

• Korean Journal of Agricultural Science
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• v.14 no.2
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• pp.413-421
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• 1987

The objective of this study was to evaluate the possibility of sex preselection by gradients methods using bovine serum albumin in rabbits. Artificial insemination with separated sperm was performed, after highly motile sperm were separated by different methods using 6%, 10% and 20% bovine serum albumin. Various characteristics of separated sperm, and the conception rate and secondary sex ratio at artificial insemination with sperm separated by different methods were compared. The results obtained were as follows. 1. The conception rate of sperm separated by bovine serum albumin gradients was higher than th at of control sperm. But secondary sex ratio was not altered by this methods. 2. The sperm separated by bovine serum albumin gradients showed significantly high value in motility, percent of normal sperm and progressive motility as compared with control sperm and revealed the highest sperm recovery when separated with 6% bovine serum albumin. 3. The sperm motility, percent of normal sperm and progressive motility of the highly motile sperm frozen after being separated from raw semen with bovine serum albumin, showed significantly high value than those of control sperm.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.3
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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.

• Reproductive and Developmental Biology
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• v.40 no.4
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• pp.33-37
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• 2016

This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.