• Clinical and Experimental Reproductive Medicine
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• v.20 no.1
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• pp.65-70
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• 1993

To maintain a good sperm motility is one of the key factors for the successful artificial insemination in retrograde ejaculation, and the sperm motility has been shown to be affected by various environmental factors, including change in pH and osmolarity. Herein we have analyzed the effect of change in pH and osmolarity in urine and normal saline on sperm motility by Sperm Quality Analyzer and Makler counting chamber. Semen, which sampled by masturbation from a 28 year old male and showed normal finding on semen analysis, was used for this study. The results were as follows: 1. When osmolarity was fixed to 300mOsm, pH did not show a definite effect on the sperm moility. However, the motility was generally a bit better in alkaline urine and saline than in acid, particularly than in pH 5.0. 2. When pH was fixed to 7.5, sperm motility was best in urine and saline of 300mOsm. Hyperosmolarity had more adverse.effect on the motility than hypoosmolarity. 3. The sperm motility was worse in the urine than in saline under the same pH and osmolarity. In conclusion, osmolarity has a definite effect on sperm motility, where as pH has relatively little effect. And certain components of urine other than pH and osmolarity might affect the sperm motility.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.9
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• pp.1199-1203
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• 2011

The objective of this study was to examine the binding potential of sperm to the epithelium of the sperm storage tubules (SST) in vitro and in vivo to assess the functional maturation of fowl sperm. Sperm from the testis, epididymis, as well as the proximal, middle and distal vas deferens were incubated in vitro with either the uterovaginal junction (UVJ)- or infundibular tissue containing SST at $39^{\circ}C$ for 30 min. Aliquots of sperm were also artificially inseminated into the uteri of hens, and the UVJ and infundibulum were collected 24 h post artificial insemination (AI). After incubation and AI, tissues were washed to remove loosely adhered sperm and subjected to fluorescence staining with 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) for counting the number of bound sperm per 0.25 mm2 of surface area. Sperm from the testis, epididymis, and the three segments of the vas deferens exhibited their differential (p<0.05) binding capacity, which increased gradually from the testicular to distal vas deferens sperm under both in vitro and in vivo conditions. Existing similar trend, sperm, regardless of their source had a lesser affinity to bind to the epithelium of the infundibular SST than to the UVJ-SST. These experimental results suggested that fowl sperm may undergo gradual changes in the process of functional maturation, whereby they gain the ability to bind to the epithelium of the SST during their passage through the male reproductive tract (MRT).

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.163-166
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• 2012

This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22~24 hr incubation from counting agar plate in which sperm dilute to $10{\sim}10^6$ in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.

• Korean Journal of Poultry Science
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• v.27 no.1
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• pp.63-71
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• 2000

The testis is an extremely heterogeneous organ, containing numerous compartments types. Morphometric studies were performed of 3 avian species (pigeon, pheasant and chicken) to determine volume density absolute volume, numerical density, total number of serminiferous tubule components, and sperm production, especially those related to the Sertoli cell, and to make comparisons among the species. Volume density of seminiferous tubule components per testis was determined by point counting method. Testis volume and sperm production were measured by routine techniques. Numerical density (the number of cells per unit volume of testis) of seminiferous tubule components per testis was determined by morphometry (Floderus method). The volume density of seminiferous tubules per testis was 91.58, 92.18 and 94.21% in pigeon, pheasant, and chicken, respectively. The volume density of spermatogonium, spermatocyte, spermatid, spermatozoon, and Sertoli cell did not produce significant changes in the three species. The absolute volume of spermatogonium, spermatocyte, spermatid, and Sertoli cell showed significant changes in the three species (p<0.05). The average volume of Sertoli cell ranged from 758.34(pheasant) to 1,212.9 ㎛$^3$(chicken) and was not significantoy different in the three species(p>0.05). The number of Sertoli cells per testis showed significant differences in the three species : 34.52 $\times$10(sup)6, 186.82$\times$10(sup)6, 810.62$\times$10(sup)6 in pigeon, pheasant, and chicken, respectively(p<0.05). The sperm production was significantly different in the three species : 3,018$\times$10(sup)6, 993.9$\times$10(sup)6, and 8.9$\times$10(sup)6 in chicken, pheasant, and pigeon, respectively(p<0.05). These results suggest that number of Sertoli cells may be more important than Sertoli cell size in explaining the difference in sperm production among the three species.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.340-345
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• 2008

Oxidative stress is one of the major causes of failure of in vitro storage of boar semen. Reactive oxygen species (ROS) are one of the important mediators of oxidative stress during in vitro storage of boar semen. Our study examined the effects of taurine on sperm characteristic and on in vitro developmental embryos during in vitro storage of boar semen for 7 days. Semen was randomly aliquoted into 3 centrifuge tubes and treated with different concentrations of taurine (25-100 mM). The characteristics of boar sperm were analyzed for motility by light microscopy, viability by using a Makler counting chamber and membrane integrity by a hypoosmotic swelling test (HOST). The percentages of motile spermatozoa in taurine groups after 5 days were significantly higher compared to the control. Sperm viability in the control was lower than in taurine groups after 7 days irrespective of different taurine concentration. In the hyoosmotic swelling test (HOST), significantly higher results were obtained in taurine groups after 3 days. Also, the developmental rates of IVM/IVF porcine embryos from semen treated with pyruvate and taurine were significantly increased when compared with the control (p<0.05). These results indicate that supplementation of taurine as an antioxidant in boar semen extender can improve the semen quality.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.61-65
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• 2011

The objective of this study was to examine the effect of various discontinuous Percoll washing conditions on motile sperm recovery rate and motion kinematics. Frozen semen samples from 3 bulls (0.5 ml plastic straws, 6% glycerol in egg yolk-Tris-glycerol extender) were thawed in $37^{\circ}C$ water bath for 1 min. After thawing, the mixed semen samples were randomly allocated to 12 treatment groups. Briefly, the spermatozoa were centrifuged for three different time lengths (10, 20, and 30 min) at two gravities ($300{\times}g$ and $700{\times}g$) through two concentrations of discontinuous Percoll density gradient of 1 ml 90%: 1 ml 45% Percoll and 2 ml 90%: 2 ml 45% Percoll to remove extender, debris, and dead spermatozoa. Motile sperm recovery rate and motion kinematics were evaluated by computer assisted sperm analyzer using Makler counting chamber. Sperm motility (%) and motile sperm recovery rate showed similar pattern in all treatment groups. However, sperm motility (%) and motile sperm recovery rate were highest at $700{\times}g$ for 30 min through a discontionous Percoll density gradient of 1 ml 90%: 1 ml 45% Percoll. There were no significant differences in motion kinematics after various Percoll washings. These results suggest that force of centrifugation, centrifugation time, and Percoll volume significantly affect motile sperm recovery rate.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.7
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• pp.847-852
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• 2006

This study was designed to investigate the effects of KH204 on the relaxation response and reproductive function in male. Strips of rabbit corpus cavernosum were prepared for mounting and isometric tension measurement in an organ bath. On cavernosal strips contracted with $1{\times}10^{-6}$ phenylephrine and KH204 was applied in increasing concentrations from 0, 61, 183 and 549 mg/L, causing dose-dependent relaxation. Male Sprague-Dawley rats were orally administered with 61, 183 and 549 mg/kg/day of KH204 for 10 weeks. We examined organ weights, testicular sperm head counts, epididymal sperm counts, motility and morphology. KH204 relaxed rabbit corpus cavernosal strip contracted by $1{\times}10^{-6}$ phenylephrine in a dose-dependent manner. In the male rat, testicular weight was increased significantly in the KH204 treated groups compared with control group. Also in the testicular sperm head counts, epididymal sperm counts were increased significantly in the KH204-treated rats. In conclusion, the data suggest that KH204 could enhance erectile and reproductive function.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.898-908
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• 1998

The aim of this experiments were to assess the time-interval change of motional characteristics in frozen-thawed semen of Korean native cattle (KNC) by using computer aided semen analysis (CASA) technology. Twenty-six KNC frozen semen straws were obtained from Korean KNC improvement department, livestock improvement main division, national livestock cooperatives federation in Korea. Specimens were allowed to thaw at $37^{\circ}C$ for 30 sec in water bath. Semen analysis was performed on semen image analysis system (SIAS, Medical supply, Korea) adjusted to the gate settings and used the semen droplet ($5{\mu}l$) placed on Makler counting chamber (Sefi medical instrument, Israel) prewarmed at $37^{\circ}C$. The same person used the same micropipette to fill the Makler counting chamber. A total of 150 or more of sperms were analysed in each specimen by a single trained person by scanning at least 5 to 10 fields. The measurement parameters in SIAS were as follows ; frame rate = 30 frames per sec, image capture = 1 sec, minimum motile speed = $10{\mu}m/s$, maximum countable sperm number = 400. Statistical analysis was done by Student t-test with use of the Sigma plot program on a IBM personal computer. The dancemean(DNM) and hyperactivated sperm(HYP) of frozen-thawed KNC semen kinematics were significantly decreased(p < 0.05) after 10 min of incubation at $37^{\circ}C$ water bath. But, wobble(WOB) of same sample semen was significantly increased(p < 0.05) after 10 min of incubation and significantly decrease(p < 0.05) after 60 min of same incubation. And, after 30 mim of incubation, significantly differences were found most of motion kinematics, motifity(MOT), curvilinear velocity(VCL), straight line velocity(VSL), average path velocity(VAP), amplitude of lateral head displacement(ALH), beat cross frequency(BCF), mean angular displacement(MAD), dance(DNC), on same sample semen. The DNM of KNC semen sample was variable kinematics after 30 min of incubation. Also, the linearity(LIN) and straightness(STR) was significantly decreased(p < 0.05) from 60 min of incubation. In conclusion, the AI within 30 min after thawing of frozen semen can be an effective method for obtaining high fertility rate in KNC reproductive program.

• Fisheries and Aquatic Sciences
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• v.20 no.11
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• pp.29.1-29.7
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• 2017

The objective of this study was to determine a simple and reliable ploidy identification protocol for the rainbow trout (RT), Oncorhynchus mykiss, in the field condition. To evaluate the ploidy level and compare different detection protocols, triploid RT and gynogenesis were induced by UV irradiation and/or heat shock. The hatching rate at day 30 was 85.2% and the survival rate at day 90 was 69.4% (fingerling). The sex ratio of female RT was 93.75% in the gynogenesis group, illustrating that the UV irradiation inactivated the sperm DNA. The hatching rate and survival rate were 82.0 and 74.7%, respectively, in the triploid-induced group. The triploid induction rate by heat shock procedure was 73.9%. Cytogenetic protocols for ploidy identification such as chromosome counting, erythrocyte nuclear size comparison, and analysis of nucleolar organizing regions (NORs) by silver staining were compared. Silver nitrate staining showed the greatest success rate (22/23 and 32/32 for the triploid-induced group and gynogenesis group, respectively), followed by erythrocyte nuclear size comparison (16/23 and 19/32 for the triploid-induced group and gynogenesis group, respectively) and, lastly, chromosome preparation (2/23 and 6/32 for the triploid-induced group and gynogenesis group, respectively) with the lowest success rate. Based on our findings, silver staining for RT ploidy identification is speculated to be highly applicable in a wide range of research conditions, due to its cost-effectiveness and simplicity compared to other numerous ploidy detection protocols.