• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.612-616
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• 2004

This study was carried out to compare the semen characteristics, frozen-thawed sperm viability and testosterone concentration and in vitro fertilization (IVF) and development of in vitro matured pig oocytes between two Yorkshire boars. Semen and blood samples were collected once per week from October to November 2002 from two adult Yorkshire boars at 18 months of age with 170 kg body weight. Sperm were deep frozen in 5 ml maxi-straws with lactose-egg yolk and N-acetyl-D-glucosamine (LEN) diluent and stored in liquid nitrogen. Blood samples were obtained at 10 a.m. by inserting a 21 gauge, hypodermic needle attached to 10 ml syringe into surface veins in the ear. The concentration of testosterone was determined by Competitive Enzyme Immunoassay. Ovaries were collected from prepubertal gilts at a local slaughter house. Cumulus oocyte complexes were aspirated from antral follicles (3 to 6 mm in diameter). The medium used for oocyte maturation was modified TCM 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at $38.5^{\circ}C$, 5% $CO_2$ in air. For IVF, one frozen 5 ml straw was thawed at $52^{\circ}C$in 40 sec and was diluted with 20 ml Beltsville thawing solution at room temperature. Sperm were washed 2 times in mTLP-PVA and inseminated without preincubation after thawing. Oocytes were inseminated with $2{\times}10^7$/ml sperm concentration. Oocytes were coincubated for 6 h in 500 ${\mu}$l mTBM fertilization medium. At 6 h after IVF, oocytes were transferred into 500 ${\mu}$l NCSU-23 culture medium for further culture of 48 and 144 h. There were no significant differences in the semen volume, motility, normal acrosome morphology and sperm concentration of raw semen between A and B of Yorkshire boar. However, motility and normal acrosome of boar A were higher than those of boar B at 0.5, 2, 3, 4, 5 and 6 h incubations of frozen-thawed sperm. Testosterone concentration (3.75 ng/ml) of boar A was higher than that (2.34 ng/ml) of boar B. The rate of blastocyst formation (15.1%) of boar A was higher than that (10.4%) of boar B. In conclusion, serum testosterone concentration of boar showed very important role for the frozen-thawed sperm viability and the blastocyst formation of pig oocytes matured in vitro.

• 한국발생생물학회지:발생과생식
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• 제17권4호
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• pp.345-351
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• 2013

This study examines the effects on fertilization rate (FR), hatching rate (HR), and normal individual rate after artificial fertilization using frozen thawed sperm according to the cryoprotectant (DMSO) concentration and the period of cryopreserved sperm of longtooth grouper, Epinephelus bruneus. Performing artificial fertilization using frozen-thawed sperm, after freezing the sperm at different DMSO concentration of 5.0%, 7.5%, 10.0% respectively, FR were (DMSO 5.0%: $99.5{\pm}0.8%$, DMSO 7.5%: $99.5{\pm}0.7%$, and DMSO 10.0%: $99.6{\pm}0.6%$). The results are not significantly different from the control fresh sperm (100%). HR also (DMSO 5.0%: $96.2{\pm}2.3%$, DMSO 7.5%: $95.3{\pm}3.6%$, 10.0%: $96.6{\pm}1.8%$) were not significantly different in each group. The normal individual rate after hatching using with control fresh sperm ($98.4%{\pm}0.5$) and DMSO concentration level of 5.0% ($97.8{\pm}0.1%$) were not significantly different. However, with 7.5% ($97.2{\pm}0.6%$) and 10.0% DMSO concentrations ($95.9{\pm}0.2%$) are lower than the normal individual rate after hatching observed in the control and 5.0% DMSO. Performing artificial fertilization using frozen-thawed sperm at different frozen period (2 days, 2 years, and 3 years), 10% DMSO FR and HR of 3 years (FR; $66.8{\pm}1.8%$, HR: $82.0{\pm}12.9%$) and 2 years (FR; $78.5{\pm}14.8%$, HR: $79.3{\pm}0.6%$) cryopreserved sperm were lower than control (FR; 100%, HR: $91.1{\pm}3.6%$) and 2 days cryopreserved sperm (FR; $99.6{\pm}0.6%$, HR: $96.6{\pm}1.8%$). These results suggest suitable DMSO concentration ranges of cryopreservation sperm for E. bruneus is 5 to 10% and with 2 to 3 years cryopreservation period, cryopreservation sperm can be useful for seed production.

• 한국가축번식학회지
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• 제22권3호
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• pp.213-219
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• 1998

This study was carried out to investigate on the improvement of fertilizing ability of in vitro matured oocytes from sperm density, motility and polyvinylpyrrolidine (PVP) concentration, by intracytoplasmic sperm injection(ICSI) into the bovine oocytes. 1. The in vitro fertilization and cleavage rates of oocytes from 1.0, 2.0, 3.0, 5.0 ($\times$106/ml) sperm concentration by IVF and ICSI of bovine oocytes were 45.0%~65.0%, 65.0%~90.0% and 10.0%~30.0%, 35.0%~70.0%, respectively. 2. The in vitro fertilization and cleavage rates of oocytes from 20, 40, 60, 80% of sperm motility by IVF and ICSI of bovine oocytes were 47.8%~75.0%, 78.3%~90.0% and 8.7%~25.0%, 34.8%~70.0%, respectively. 3. The in vitro fertilization and cleavage rates of oocytes from 0.01, 0.02, 0.03, 0.05% of PVP concentration by microinjection of single into the bovine oocytes were 72.7%, 90.9%, 83.3%, 76.9% and 45.5%, 72.7%, 58.3%, 61.5%, respectively and these values of 0.02% addition of PVP were higher than other concentrations of PVP. 4. The in vitro fertilization and developmental rates of oocytes by IVF and ICSI methods were 63.3%~64.6%, 26.7%~29.2% and 88.2%, 47.1%, respectively. This ICSI method was improved high fertilization rates of bovined oocytes.

• Asian-Australasian Journal of Animal Sciences
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• 제21권8호
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• pp.1127-1133
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• 2008

This study was designed to develop a method of liquid storage of boar sperm at $4^{\circ}C$ by using the modified Beltsville F5 (BF5) diluent with bovine serum albumin (BSA) and N-acetyl-D-glucosamine. Boar sperm were stored in lactose-egg yolk and N-acetyl-D-glucosamine (LEN), BF5 and Golden-Pig liquid 4 (GPL4) diluents at $4^{\circ}C$ for 5 days and were examined for sperm viability, adenosine triphosphate (ATP) and in vitro fertilization (IVF). The percentage of sperm viability in GPL4 diluent was higher than in LEN and BF5 diluent from 1 to 5 days of storage at $4^{\circ}C$. The percentage of sperm viability steadily declined from 1 to 5 days of storage in the three different diluents. Sperm ATP in GPL4 diluent was higher than in LEN and BF5 diluents from 1 to 5 days of storage. Sperm ATP rapidly declined after 5 days of storage in the three different diluents. Porcine oocytes matured in vitro were inseminated with different sperm concentrations of liquid semen stored for 3 days in GPL4 diluent. The percentage of monospermic oocytes did not show any differences from 2.5 to $20{\times}10^5$ sperm/ml. However, the percentage of polyspermic oocytes in the sperm concentration of $2.5{\times}10^5$ sperm/ml was lower than in concentrations of 5, 10 and $20{\times}10^5$5 sperm/ml. The percentage of blastocysts from the cleaved oocytes at $2.5{\times}10^5/ml$ sperm concentration was significantly lower than at 5, 10 and $20{\times}10^5sperm/ml$ concentrations. In conclusion, GPL4 diluent can be stored at $4^{\circ}C$ for 5 days and showed higher sperm viability and sperm ATP concentration compared with LEN and BF5 diluents. Also, we found that GPL4 diluent can be used for IVF of porcine oocytes.

• 대한한의학회지
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• 제27권3호
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• pp.63-76
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• 2006

Objective: These studies were undertaken to evaluate the effects of the different administration duration of Morindae Radix extract solution on spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testes and the activities of sperm hyaluronidase. Materials and Method: We used 8-week-old ICR mice and administered 0.3mg/g extract solution of Morindae Radix once a day for 30, 60, 90 and 120 days. The control group was administered normal saline in the same way and duration. We examined the number of total, motile and normal sperm from the cauda epididymis. We also compared the testicular tissue, especially seminiferous tubules, between the control and treated groups by histochemical methods. Finally, we observed the difference of sperm hyaluronidase activities between the control and treated groups. Results: Significant administration duration-dependent differences were observed in the concentration of total sperm, motility and normality of spermatozoa of the Morindae Radix extract solution administered groups compared to the control group. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Morindae Radix extract solution administered groups compared to the control group. Also, the activity of hyaluronidase was significantly increased in the Morindae Radix extract solution administered groups compared to the control group. Conclusions: This study shows that the beneficial effect of Morindae Radix extract solution on the concentration, motility and morphology of sperm, the testicular tissues and the activities of sperm hyaluronidase increased the greater the duration the mice were administered it. We suggest that Morindae Radix may be useful for the treatment of male sexual dysfunction and infertility.

• Animal Bioscience
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• 제36권11호
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• pp.1647-1654
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• 2023

Objective: Aging roosters typically exhibit subfertility with decreasing semen quality, furthermore Thai native roosters reared in rural areas are raised for a longer duration than their usual lifespan. The present study therefore aimed to assess the effect of selenium supplementation as an antioxidative substance in diets to improve the semen cryopreservation of aged roosters. Methods: Semen samples were collected from young (n = 20) and aged (n = 20) Thai native roosters (Pradu Hang Dum) at 36 and 105 weeks of age when starting the experiment, respectively. They were fed diets either non-supplemented or supplemented with selenium (0.75 ppm). Fresh semen quality and lipid peroxidation of fresh semen was evaluated before cryopreservation using the traditional liquid nitrogen vapor method. Post-thaw sperm quality and fertility potential were determined. Results: Advancing age is unrelated to decreasing fresh semen quality (p>0.05). However, lipid peroxidation in rooster semen depended on age, and the malondialdehyde (MDA) concentration increased in aged roosters (p<0.05). Selenium supplementation in diets significantly decreased the MDA concentration and increased the sperm concentration (p<0.05). In contrast, cryopreserved semen was affected by advancing rooster age, and selenium influenced sperm quality (p<0.05). Younger roosters had higher post-thaw sperm quality and fertility potential than aged roosters (p<0.05). Likewise, diet selenium supplements improved post-thaw sperm quality and fertility compared with the non-supplement group. Conclusion: Rooster's age does not influence the rooster sperm quality of fresh semen, while sperm cryotolerance and fertility were greater in young roosters than in aged roosters. However, sperm of aged roosters could be improved by dietary selenium supplementation.

• Clinical and Experimental Reproductive Medicine
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• 제51권1호
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• pp.48-56
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• 2024

Objective: This study investigated the relationship of anthropometric and metabolic risk factors with seminal and sex steroidal hormone parameters in a screened population of healthy males. Methods: The participants were healthy young men without chronic or congenital diseases. The body composition parameters that we investigated were measured weight, height, and waist circumference (WC), as well as bioelectrical impedance analysis. Semen samples were analyzed for semen volume, sperm concentration, sperm motility and morphology, seminal pH, and liquefaction time. Biochemistry analysis, including glucose and lipid metabolism parameters, was conducted on fasting blood samples. Testicular volume was calculated separately for each testis using ultrasonography. Results: Body mass index exhibited an inverse association with total sperm count. WC showed negative correlations with numerous seminal parameters, including sperm concentration, total sperm count, sperm morphology, and follicle-stimulating hormone levels. The basal metabolic rate was associated with seminal pH, liquefaction time, and sperm motility. WC, fat mass percentage, and triglyceride levels exhibited negative correlations with sex hormone binding globulin. The measures of glucose metabolism were associated with a greater number of seminal parameters than the measures of cholesterol metabolism. C-reactive protein levels were inversely associated with sperm concentration and total sperm count. Conclusion: Anthropometric and metabolic risk factors were found to predict semen quality and alterations in sex steroidal hormone levels.

• 한국수산과학회지
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• 제56권5호
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• pp.644-650
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• 2023

We examined the effects of serotonin on the spawning response, sperm motility, and D-shaped larva production in Eastern Gooeyduck clam Panopea sp. based on the sperm densities at fertilization and washing after mixing the eggs and sperm. The highest spawning induction was found showed in females and males injected with 1 mL of 2 mM serotonin. The spawning responses in females and males were higher at concentrations greater than 1 mM and 0.75 mM, respectively. Regarding the activities of sperm in sea water after serotonin injection, the sperm showed activity at >90.0% until 120 mins. We also examined the effects of sperm concentration at the fertilization and washing times after mixing the eggs and sperm. We confirmed that washing within 1 minute at a concentration of 1,500 sperms/mL or less can prevent egg destruction by polyspermy and secure a large number of D-phase larvae. These results should be useful for developing the aquaculture process for Eastern Gooeyduck clam, Panopea sp.

• Clinical and Experimental Reproductive Medicine
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• 제30권2호
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• pp.165-169
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• 2003

Objective: To evaluate the results of CASA systems and to compare its results. Methods: Fifty semen sampales were analysed. Concentration, motility and forward progression were evaluated simultaneously on the same semen samples using Cell Soft System-3000 (CS system) and Sperm Quality Analyzer-V (SQA system). Results: Mean semen volume was $2.8{\pm}1.2\;ml$. Mean value of sperm concentration, motility, forward progression using CS system were $83.4{\pm}45.7{\times}10^6/ml$, $52.3{\pm}16.4%$ and $48.6{\pm}13.4%$, respectively. And mean value of sperm concentration, motility, forward progression using SQA system were $78.2{\pm}42.9{\times}10^6/ml$, $57.0{\pm}24.0%$ and $50.6{\pm}21.9%$, respectively. There were no statistical significancy of sperm concentration, motility, forward progression between the two devices. Conclusion: SQA system variables well correlated with the CS system. As a screening test for semen quality, CS system and SQA system is considered as useful in the management of male infertility.

• 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.