Damaged spermatozoa are supposed to be trapped in glass wool. In the respect of this, two glass wool filtration spermatozoa groups (0.5 cm, 1 cm depth) were compared with control group to assay sperm motility, HOS values, and vital rate by CFDA/PI staining method following glass wool filtration. The motility of canine sperm extended with PBS+PVP after glass wool filtration was lower in both filtrated groups than that of the control group (p<0.01) and the same significant difference was also shown in canine semen extended with Tris buffer (p<0.01). The motility of canine sperm diluted with PBS+PVP was higher than that diluted with Tris buffer in the same experimental groups (p<0.05). The motility of control group was not significantly decreased until 2 hours immediately after extending, however, the motility of both glass wool filtrated spermatozoa were significantly decreased as time passed until 2 hours after filtration (p<0.01). At each time for assay (immediately, 30 min, 2 hours after filtration), the motility of canine sperm of control group was higher than the filtrated groups (p<0.05), whereas the motility of 0.5 cm depth group was higher than 1 cm depth group at the immediate time after filtration (p<0.05), 30 minutes later (p<0.05) with no difference at 2 hours. No difference was shown among the experimental groups in HOS values of canine sperm after glass wool filtration. The vital rate assayed by CFDA/PI staining of both filter groups was higher than the control group (p<0.05).
Baek, Sun Young;Chung, Hak Jae;Hong, Joon Ki;Cho, Eun Seok;Choi, Inchul
Korean Journal of Agricultural Science
/
v.47
no.3
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pp.657-665
/
2020
The objective of this study was to investigate whether supplementation of pentoxifylline (PTX; phosphodiesterase inhibitor) to thawed boar semen improves the post-thaw motility of sperm and affects the efficiency of artificial insemination (AI) and further development. To determine the concentration of PTX for AI, frozen-thawed semen was incubated with 0, 5, 10, and 20 mM PTX in an extender freezing medium, respectively, after thawing. Kinematic properties of sperm were examined with a computer-assisted semen analysis (CASA) system. In addition, viability and mitochondrial activity were also tested by LIVE/DEAD and a MitoTracker kit. There were no significant differences in the kinetic parameters of thawed sperm between control and treatment groups, but overall assessment parameters such as motility and rapid progressive were higher in the 10 mM PTX group. In the viability and mitochondrial assay, there were no significant differences observed in the PTX treatment, compared to the control. For further analysis, artificial inseminations were performed using frozen semen and 10 mM PTX treated cryopreserved semen, respectively. There were no differences in pregnancy rates and fetus weights among the groups until 30 and 40 days, but litter size was reduced and relatively low-birth weight was observed in the PTX group. In summary, our findings suggest that enhancement of in vitro sperm quality or non-toxicity supplemented by PTX may have detrimental effects on fetus development.
Sa, Soo-Jin;Kim, In-Cheul;Choi, Sun-Ho;Hong, Joon-Ki;Kim, Du-Wan;Cho, Kyu-Ho;Kim, Young-Hwa;Chung, Ki-Hwa;Park, Jun-Cheol
Journal of Embryo Transfer
/
v.28
no.4
/
pp.349-353
/
2013
The objective of this study was to compare the effect of semen extenders on the sperm motility, viability, acrosome integrity and functional integrity of plasma membrane (HOST: hypo-osmotic swelling test) during liquid preservation of Korean Native boar semen. In this experiment, semen was diluted in Androhep plus, Beltsville Thawing Solution (BTS), ModenaTM, Seminark and Vitasem LD. Sperm-rich fractions were collected from three Korean Native boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different semen extenders. Semen samples were stored at $17^{\circ}C$ for 96 hours. On everyday (0, 24, 48, 72, 96 h) after storage, the sperm characteristics relevant for fertility, such as sperm motility, viability, acrosome integrity and HOST positive were evaluated. The motility of spermatozoa stored in different extenders was no significantly different among other extenders (P>0.05). Also, no difference was observed among samples processed with different extenders in the percentage of sperm viability, acrosome integrity and HOST positive. All extenders maintained a high percentage (70%) of sperm motility, viability and acrosome integrity through 96 h of storage. The result of this study show that there was no significant differences among extenders in their capacity to preserve motility, viability and membrane integrity of spermatozoa from normal, fertile Korean Native boars for 96 h of liquid preservation at $17^{\circ}C$.
The objective of this study was to investigate the effect of bacterial contamination on elapsed time after preservation on boar semen. Known numbers of Escherichia coli (E. coli) were inoculated to freshly ejaculated semen and sperm parameters such as viability, motility, agglutination, acrosome integrity and hypo-osmotic swelling test were performed during 7 days of liquid preservation. Semen samples were prepared using antibiotic free BTS extender and 4 different levels of E. coli were treated to semen with following concentrations; 3,000, 5,000, 7,000, 10,000 CFU/ml of sperms. Semen samples were preserved at $17^{\circ}C$ for 7 days in semen storage until analyzed. Aliquots were subjected to measure the sperm viability, motility and agglutination using computer assisted sperm analysis (CASA) system, acrosome integrity was performed using chlortetracycline (CTC) staining method and hypo-osmotic swelling test was performed using hypotonic solution from day 1 (day of semen collection) to 7. Detrimental effects on sperm motility and viability were observed 3 days after preservation at the level of 5,000 CFU/ml (p<0.05). Percentage of sperm abnormality was higher (p<0.05) in over 5,000 CFU/ml groups. Sperm agglutination rate was also significantly higher (p<0.05) in groups of 5,000 and 7,000 CFU/ml. The rate of acrosome reacted sperm was higher as preservation time goes in all the samples but the pattern was clearly higher among E. coli contaminated groups (p<0.05). The sperm membrane integrity in terms of hypo-osmotic test, E. coli affects little compared to other sperm parameters. The deleterious effects observed due to the bacterial contamination in semen suggest that importance of hygiene protocol to minimize the bacterial contamination during semen collection and processing.
Kim Heui-Eun;Lee Jung-won;Kim Nam-Soo;Choi In-Hyuk
Journal of Veterinary Clinics
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v.10
no.1
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pp.11-18
/
1993
Four extenders such as tris-fructose-citrate, Tris-glucose-citrate, glycine-glucose-citrate and lactose that the more frequently utilized types of semen extenders used for freezing dog semen evaluated with sperm motility, viability and acrosomal score in the processing procedures to prior freezing and after frozen-thawing respectively. Each extender contained 4% glycerol and 20% egg yolk were treated by same methods in dilution, freezing, storage and thawing. The results were obtained as follows; 1. The sperm motility and viability in procedure from dilution to frozen-thawing appeared superiorly with recovery rate of 53.2%, 54.8% in tris-fructose-citrate but appeared inferiorly with recover rate of 8.4%, 8.3% in lactose to others. 2. In the processing procedure course to prior-freezing, glycine-glucose-citrate appeared superiorly with decrease rate of 5.4% in motility, and lactose with decrease rate of 4.6% in viability, but tris-glucose-citrate appeared inferiorly with decrease rate of 12.2%, 11.4% in the sperm motility and viability. 3. During frozen-thawing, tris-fructose-citrate appeared superiorly with decrease rate of 35.2% in motility and 30.7% in viability but lactose appeared inferiorly with decrease rate of 76.7% in motility and 75.7% in viability. 4. The variation of acrosome morphology in the total processing procedures appeared that glycine-glucose citrate were superior with acrosome score of 0.1191$\pm$0.029, that tris-fructose-citrate were inferior with acrosome score of 0.1941$\pm$0.045 to others.
The purpose of this study was to identify for extenders, adhesives, colorants, and fungicides suitable for the production of high-quality film-coated seeds of Chinese cabbage. In addition, the effect of various additives used for film coating on the germination of coated seeds and seedling vigor was investigated to establish appropriate treatment conditions for film coating efficiency. The germination rate and germination speed in Chinese cabbage film-coated seeds did not differ significantly depending on the type and concentration of the extender, while the use of Talc resulted in superior granulation capacity and adhesion of the seed coat in comparison to Zeolite. There was no significant difference in the germination rate depending on the type of adhesive used for film coating of Chinese cabbage seeds, but the germination rate was slightly slower in the film-coated seeds using carboxymethyl cellulose as an adhesive. On the other hand, the seeds coated with a film using 1% PVA as an adhesive not only maintained hardness enough to withstand external pressure, but also showed a high germination rate. In addition, the suitable disinfectant for film coating was venoram, and the treatment concentration was good at 250 mg·L-1. The type of colorant used in the film-coated seeds of Chinese cabbage did not significantly affect the germination rate, germination speed, and seedling vigor. The seeds film-coated using the liquid colorant were superior in appearance to the seeds film-coated with the solid colorant, which implies the possibility of reduction in the manufacturing costs by using the water-soluble paints as colorants for film-coated seeds.
Objective: This study investigated the effect of adding seminal plasma to frozen-thawed semen on the quality of sperm and pregnancy following insemination in dromedary camels. Methods: In experiment 1, the frozen-thawed semen from 9 collections (3 bulls) was further diluted with either the base extender or homologous seminal plasma (HSP). In the second experiment, a pooled sample of frozen-thawed semen was diluted with either seminal plasma from another three bulls. Live percentage, total and progressive motility, functional and acrosome integrity, and sperm kinematics were evaluated at 15, 60, and 120 minutes post-thawing and compared to the non-treated control. In experiment 3, frozen semen was used to inseminate camels in the following experimental groups: 1-Single insemination with double dose undiluted frozen semen (n = 9); 2-Re-insemination in 6 hours with undiluted semen (n = 13); 3-Single insemination with HSP treated sperm (n = 14). Results: Frozen-thawed sperm diluted in HSP or the non-homologous seminal plasma from Bull C indicated an improvement in all parameters after 1 hour post-thawing incubation (p<0.05). The proportion of total and progressively motile sperm did not drop significantly at 60 minutes post-thawing when diluted with the seminal plasma of Bull C (p>0.05). Double insemination with nontreated sperm and single insemination with HSP-treated sperm resulted in similar pregnancy rates (15.3% vs 21.4%, p>0.05). None of the camels conceived with double-dose single insemination of nontreated sperm. Conclusion: Seminal plasma improves sperm longevity and motility after thawing in dromedary camel with a significant between-bull variation in effect. Low post-thaw sperm longevity might be the cause behind the low pregnancy rates in frozen semen insemination of dromedary camels.
Zhendong Zhu;Haolong Zhao;Qitai Yang;Yajing Li;Ruyuan Wang;Adedeji Olufemi Adetunji;Lingjiang Min
Animal Bioscience
/
v.37
no.5
/
pp.852-861
/
2024
Objective: The present study aimed to investigate the effect of β-nicotinamide mononucleotide (NMN) supplementation on ram sperm quality during storage at 4℃ in vitro. Methods: Tris-citric acid-glucose solution containing different doses of NMN (0, 30, 60, 90, and 120 µM) was used to dilute semen collected from rams and it was stored at 4℃. Sperm motility, plasma membrane integrity as well as acrosome integrity were evaluated at 0, 24, and 48 h time points after storage at 4℃. In addition, sperm mitochondrial activity, lipid peroxidation (LPO), malondialdehyde (MDA) content, reactive oxygen species (ROS) content, glutathione (GSH) content, superoxide dismutase (SOD) activity, and apoptosis were measured at 48 h time point after storage at 4℃. Results: Results demonstrate that the values obtained for sperm motility, acrosome integrity, and plasma membrane integrity in the NMN treatments were significantly higher than control (p<0.05). The addition of 60 µM NMN significantly improved ram sperm mitochondrial activity and reduced LPO, MDA content, and ROS content compared to control (p<0.05). Interestingly, sperm GSH content and SOD activity for the 60 µM NMN treatment were much higher than those observed for control. NMN treatment also decreased the level of Cleaved-Caspase 3, Cleaved-Caspase 9, and Bax while increasing Bcl-2 level in sperm at 48 h time point after storage at 4℃. Conclusion: Ram sperm quality can be maintained during storage at 4℃ with the addition of NMN at 60 µM to the semen extender. NMN also reduces oxidative stress and apoptosis. Overall, these findings suggest that NMN is efficient in improving the viability of ram sperm during storage at 4℃ in vitro.
Background: Using cryovial for freezing dog spermatozoa provides a practical method to increase extended sperm volume and shorten the time required for equilibration by using a simple freezing techniques. The purpose of this study was to determine the optimal thawing condition for dog sperm cryopreservation using cryovials. Methods: For sperm freezing, cryovials with 200 × 106 sperm/mL were cooled after the addition of tris egg yolk extender (TEY) at 4℃ for 20 min, then TEY with 4% glycerol was added and equilibrated for another 20 min before being aligned over LN2 vapor for another 20 min and plunged directly into LN2. Spermatozoa were thawed in a water bath at 37℃ for varying times (25 sec, 60 sec, 90 sec, and 120 sec) in the first experiment. In the second experiment, spermatozoa were thawed in a water bath at various temperatures and times (37℃ for 1 min, 37℃ for 1 min with gentle stirring, 24℃ for 24 min, and 75℃ for 20 sec). In these experiments, the effect of thawing conditions on motility parameters, viability (SYBR-14/PI), and acrosome integrity (PSA/FITC) of spermatozoa were investigated. Results: The post-thaw sperm motility parameters, viability, and acrosome integrity were not significantly different across the experimental groups. Conclusions: In this study, the characteristics of spermatozoa frozen using cryovials were not significantly affected by various thawing conditions.
Journal of the Korean BIBLIA Society for library and Information Science
/
v.35
no.3
/
pp.137-159
/
2024
This study analyzed the number of electronic journal subscriptions, subscription fees, usage, cost-effectiveness, etc. of university libraries to identify limitations and problems. Based on this, a case library was selected and the simulation tools Unsub was used to compare the efficiency of package subscriptions and individual subscriptions and suggest improvement measures. In the case of C University Library, SAGE and Emerald were found to be cost-effective as package subscriptions, while ScienceDirect and OUP were found to be worth considering for individual subscriptions. In particular, OUP was found to save £6,063 per year with individual subscriptions, and the ratio of accessible and usable articles reached 85.8%. In addition, ScienceDirect was found to be the most effective model for combining individual subscriptions and pay-per-view systems, considering budget savings and ease of use. Based on the results of this case study, the limitations of Unsub's application and policy tasks for developing a 'K-Unsub' version suitable for the Korean situation were suggested.
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