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.
Kim, Beom-Gi;Ham, Hyung-Bin;Kim, Sang-Hyeon;Son, Jung-Ho;Chung, Ki-Hwa
Journal of Life Science
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v.30
no.1
/
pp.77-81
/
2020
The aim of this study was to overcome some of the limiting factors that the maxi cryopreservation straw of 5 ml presents in processing boar semen. Cryopreservation of semen samples was conducted in 0.5 ml and 5.0 ml straws at two freezing rates: -140℃ in 8 minutes and 30 seconds (FR-1) and -140℃ in 14 minutes (FR-2). The straws were then thawed and the semen parameters were compared by Computer Assisted Sperm Analysis, and sperm morphology and acrosome status were examined by Coomassie blue staining. The effects of different thawing temperatures and durations were also compared, namely 37℃ for 115 sec, 50℃ for 45 sec, or 70℃ for 25 sec. In general, the FR-1 group showed higher (p<0.05) sperm viability and motility than the FR-2 group in the 5.0 ml straws. Compared to other ranges, thawing at 50℃ for 45 sec showed the highest sperm viability and motility (68.4±3.6% and 69.5±2.2%, p<0.05), suggesting that thawing temperature should be adjusted concurrently with freezing rate. Sperm morphology and acrosome integrity did not significantly differ among the groups (p>0.05). The data obtained in this study suggest that improving the freezing-thawing protocol for one artificial insemination dose straws (5.0 ml) retains the sperm's parameters from 0.5 ml cryopreservation, and is more convenient to handle, which could result in enhanced reproductive performance.
The core price policy of on-line game marketing are FPP(Fixed Pre Paid model and PPU(Pay Per Use) model. These two models have been a on-line game company's billing system and a fundamental of MMORPG in Korea. However, they took root billing system only for first movers recently. In now, the market share of several first movers is exceeding 80%, late movers witch have same billing system cannot take part in pair competition. Even though in MMORPG, many games of late movers were favorably noticed by a lot of gamers during Evaluation. Test, a lot of companies are bankrupt before make business. Late Movers declare free game first thing, they maintain their existence and win over customers in on-line game market. And next, they guarantee item selling, give multiple experience value and game money, at last, induce their customers to pay service. As it makes trouble between pay user and free user, and it linked up with the collapse of game contents balance that designed for FPP billing system, And then meet unexpected result which reduction of game life cycle. In this Paper, we classified several contents services based on game contents, and suggested contents premium services which adopted low cost strategy lead to micro payment. we hope it will apply to late movers' new billing system in MMORPG.
This study was conducted to investigate the effect of infusion of sterile boar and their semen as pregnancy antigen factor for improvement of production in gilts. We observed 160 gilts between 7 to 8 months of age that were divided into 4 treatment groups vie. A group-epidimetomized boar, B group-no sperm semen infusion, C group-vasectomized boar, and D group-control in a completely randomized design. The number of total pigs born(NT) of A, B, C, and D groups were 10.05, 10.44, 11.63, and 9.97 pigs, respectively(P<0.05). And the NT of C g.cup was the highest among treatment. The number of live pigs born(NB) was similar to NT The NB of C group (10.70) was higher than that of A(9.12) and control(9.11)(P<0.05). However, there was not significant between B and C groups. The progesterone concentration of C group was the highest compare to other group at the 6th day after breeding. However, the progesterone concentration of C was lower than other groups after 8th day. There were not significant among cortisol of A, B, C and D groups. According of the results of this study, the infusion with vasectomized boar and semen at estrus before gestation can be improved reproductive efficiency because of more litter size in gilts.
This study was conducted to determine the effect of extract of Cervi pantotrichum cornu on human sperm motility, Four different types of media were prepared such as plain Ham's F-10 medium(control medium), control medium containing 0.3% bovine serum albumin(BSA)(medium A), control medium containing the extract of Cervi pantotrichum cornu aqua-acupuncture medium(medium B) and medium B containing 0.3% BSA(medium C). Human semen were washed and divided into 4 fractions and sperm were cultured in those medium for up to 72 hours at 37$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$ in air. A total twenty eight semen samples including 14 normozoospermia and 14 asthenospermia were used for this study. In normozoospermia group, motility of control medium and medium A, B and C were 4.1%, 1.3%, 64.5%, and 77.1%, respectively after 24 hours of incubation, and were 0.0%, 0.0%. 8.8% and 44.9%, respectively after 48 hours of incubation. In asthenospermia group, motility of control medium and medium A, B and C were 2.0%, 2.2%, 58.3% and 85.1%, respectively after 24 hours of incubation, and decreased to 0.0%, 0.2%, 5.8% and 29.6%, respectively after 48 hours of incubation. In both groups, highest sperm motility was observed in medium C group when compared with other media. Furthermore motile sperm were found in medium C after 72 hours of incubation while no motile sperm was observed in the other media. Therefore it could be concluded that the extract of Cervi pantotrichum rornu affects on the human sperm motility.
Ha, A-Na;Yoon, Jin-Ho;Kim, Yu-Gon;Jo, A-Na;Lee, Kyeong-Rim;Kong, Il-Keun
Reproductive and Developmental Biology
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v.35
no.1
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pp.55-60
/
2011
The objective of this study was carried out to evaluate the efficiency of sperm collection methods on the post-thaw viability of cat semen. The cat semen was collected by artificial virginal (AV) and electronic ejaculate (EE) methods. The composition of semen extender was consisted of Tris-buffer supplemented with 20% egg yolk and 1% P/S antibiotics in Ext I, and more added 8% glycerol, 1.0% Equex STM paste of total volume in Ext II. The collected semen was adjusted the concentration and then diluted in Ext I for optimal concentration. The diluted semen was cooling to $5^{\circ}C$ temperature in refrigerator for at least 2 hrs and then diluted stepwise with Ext II for at least 1 hrs. After an equilibration for 1 hrs, the cooled semen was packaged in 0.5 ml straw and then freezing on the $LN_2$ vapor over 5 cm above from $LN_2$ and then immersed directly in $LN_2$ for cryopreservation. The frozen semen was thawed in $38^{\circ}C$ water for 15 sec and then evaluated the motility, viability, and morphology. Post-thaw semen were calculated the motility by SMI (sperm motility index). The live-dead sperm was evaluated by Eosin-B and morphological evaluation was by Diff-quik kit staining. The post-thaw concentration ($89{\times}10^6$ /ml vs. $128{\times}10^6$ /ml), viability ($22.6{\pm}10.6%$ vs. $37.1{\pm}26.1%$), morphological normality ($27.0{\pm}50.2%$ vs. $45.6{\pm}123.0%$) of EE and AV groups were not significant different, but the post-thaw motility was significant lower in EE than that in AV group ($53.1{\pm}3.6$ vs. $73.6{\pm}5.7$) (p<0.05). In conclusion, semen collection methods did not significant different between EE and AV groups except of post-thaw motility and so both semen collection methods could be applied in feline semen collection methods.
The purpose of this study was undertaken to evaluate of cryopreservation efficiency in ${\alpha}$ 1,3-galactosyltransferase knock-out(GalT KO) cloned miniature pig sperm. To compare ability of frozen-thawed sperm characteristics, three different pig strains (GalT KO) cloned miniature pig, PWG miniature pig and Duroc were used. The ejaculated semen from the three pig species was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. After thawing, the sperm ability were assessed for viability (SYBR-14/PI staining), abnormality (Rose Bengal staining), and acrosome status (intactness, intensity and capacitation) (chlorotetracycline, CTC staining). The viability of frozen-thawed GalT KO pig sperm had no significant difference as compared with Duroc and PWG miniature pig sperm. However, The CTC pattern of frozen-thawed GalT KO cloned miniature pig spermatozoa showed significantly lower rates in F pattern and AR pattern (p<0.05) and significantly higher rates in B pattern than Duroc and PWG miniature pig (p<0.05). The abnormality of GalT KO cloned miniature pig sperm was significantly lower as compared to Duroc and PWG miniature pig sperm (p<0.05). In conclusion, GalT KO cloned miniature pig semen can be cryopreserved successfully and used for artificial insemination reasonably.
In the construction project, the design field has a crucial effect on the success or failure of the final product, and it is becoming more important as a core area of national industrial development because it creates high value added by technology. In addition, since the design work is carried out based on professional technical personnel, in order to secure the quality and competitiveness of the design, it is necessary to continuously input the excellent manpower and develop the technology, and above all, the appropriate design cost must be ensured. However, as a result of reviewing the current status of the design cost criterion in Korea, problems such as mixed cost calculation method, inaccuracy of construction cost ratio method, and inactivation of cost plus fixed fee method were analyzed. Therefore, it is difficult to calculate the fair value reflecting the characteristics of the project when calculating the construction design price. Therefore, the price criterion should be improved so that the fair value of the design can be determined according to the contents of the project, By analyzing the correlation and multiple regression analysis of the projects and price information of the completed road construction design, the factors influencing the design price and the level of influence were analyzed.
Objectives: To determinate the optimal culture condition to maintain lifespan in human sperm, we evaluated the effect of different temperature on sperm motility and viability up to 5 days in normal specimens. Methods: Ejaculated semen samples with normal semen parameters were gently washed in HEPES buffered Tyrod's-Albumin-Lactate-Pyruvate (HTALP) media. Each 5 ml of HTALP + 0.3% albumin with $1{\times}10^6$ sperm/ml was incubated for 5 days in $37^{\circ}C$, $22^{\circ}C$, and $4^{\circ}C$. The sperm motility and kinematics were analyzed using computer assisted sperm analysis (CASA), membrane integrity was assessed by hypoosmotic swelling test (HOST), and capacitation status was evaluated by chlorotetracycline (CTC) fluorescence pattern. Each parameter was measured on day 1, 3, and 5, respectively. Results: The motility, viability and live/uncapacitated pattern were demonstrated significantly in temperature- and time-dependent difference (p<0.05). While the sperm cultured for 1 day in each temperature was not significantly different, the sperm cell kept in $22^{\circ}C$ after 3 days were preserved sperm motility, viability, membrane integrity, and F pattern better than in other culture temperatures. Conclusions: HTALP can be used a basic medium for culture and longevity preservation, and sperm cell kept at $22^{\circ}C$ is beneficial for assisted reproductive techniques.
This study was conducted to evaluate the feasibility of in situ soil flushing for TPH-contaminated soil remediation with column test. The soil texture of the soil was sand and the initial TPH concentration was $9,369mg\; kg^{-1}$. 0.1% Tween-80 was selected as surfactant solution. And the acrylic and the glass syringe columns were used as reactors. In the acrylic column test, 35% of the initial TPH was removed in 1 PV of flushing and approximately 40% in 5 PV and finally 7 PV showed about 60%. The glass column test showed 3 ~ 12% higher removal efficiency than that of acrylic test until 5 PV of flushing. However, there was no difference in TPH removal efficiency when 7 PV of surfactant was finally flushed. Both of alum only and alum+polymer mixed surfactants showed also the best coagulation efficiency in $150mg\;L^{-1}$ of concentraion. When Tween 80 was newly dissolved in 0.1% to the recovered solution after the coagulation treatment, the removal efficiency was increased from 32.0% to 41.0% in comparison to the new 0.1% Tween 80 solution without reuse by coagulation treatment.
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