• Mycobiology
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• v.44 no.1
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• pp.54-57
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• 2016

Davallialactone (DAVA) is a hispidin analogue derived from the medicinal fungus Phellinus baumii. We examined the effect of DAVA on in vitro fertilization (IVF) of pigs. Boar spermatozoa were incubated in fertilization medium with varying concentrations of DAVA, then sperm motility and reactive oxygen species (ROS) level were evaluated. Higher sperm motility was found following the addition of 0.5 or $1{\mu}M$ DAVA after incubation than addition of other concentrations or controls. ROS level decreased significantly with the addition of DAVA. The rate of normal fertilization was higher in the presence of $1{\mu}M$ DAVA (65.1%) than were those of other concentrations or controls (45.4~59.4%), and the highest total fertilization rate (mono- and polyspermic oocytes) was observed at $1{\mu}M$ DAVA (83%). In conclusion, addition of DAVA to fertilization medium improved sperm motility, and reduced ROS level so as to potentially improve sperm-oocyte binding in IVF, suggesting the potential of a compound isolated from mushrooms in assisted reproductive technology for humans and animals.

• Journal of Animal Science and Technology
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• v.64 no.6
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• pp.1024-1034
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• 2022

Identifying effective biomarkers for the diagnosis of male fertility is crucial for improving animal production and treating male infertility in humans. Ras-related proteins (Rab) are associated with morphological and motion kinematic functions in spermatozoa. Moreover, Rab2A, a Rab protein, is a possible male fertility-related biomarker. The present study was designed to identify additional fertility-related biomarkers among the various Rab proteins. First, the expression of Rab proteins (Rab3A, 4, 5, 8A, 9, 14, 25, 27A, and 34A) from 31 duroc boar spermatozoa was measured before and after capacitation; correlation between Rab protein expression and litter size was evaluated by statistical analysis. The results showed that the expression of Rab3A, 4, 5, 8A, 9, and 25 before capacitation and Rab3A, 4, 5, 8A, 9, and 14 after capacitation were negatively correlated with litter size. Moreover, depending on the cutoff values calculated by receiver operating curves, an increase in litter size was observed when evaluating the ability of the Rab proteins to forecast litter size. Therefore, we suggest that Rab proteins may be potential fertility-related biomarkers that could help select superior sires in the livestock industry.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.317-325
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• 2001

This study investigated the effect of ferrous sulfate (Fe$^{2+}$) and/or ascorbic acid (Asc) on fertilizing ability in vitro of frozen-thawed boar spermatozoa. Using chlortetracycline (CTC) fluorescence, the spermatozoa was treated in preincubation medium with control, Fe$^{2+}$(1 mM), Asc (0.5 mM) and Fe$^{2+}$Asc to assessed for acrosome reaction, and the oocyte penetration test to determine whether the Fe$^{2+}$ and/or Asc can promote the penetration ability in vitro. When frozen-thawed spermatozoa was washed with preincubation medium, there were significantly (P ＜ 0.05) more acrosome-reacted in medium with Fe$^{2+}$Asc (38%) than control (27%). The penetration rates were also significantly (P ＜ 0.05) higher in medium with Fe$^{2+}$Asc (76%) than control (55%). Next, the lipid peroxidation of sperm was evaluated on the basis of malondialdehyde production following same treatments. The addition of Fe$^{2+}$Asc to sperm suspension increases the formation of malondialdehyde. However, there were not significantly different under the all conditions. The sperm suspension were also treated with control, Fe$^{2+}$, Asc and Fe$^{2+}$/Asc and assayed for sulfhydry1(-SH) group content. In the Fe$^{2+}$/Asc group, sperm-SH group were higher than another groups. In spermatozoa treated with Fe$^{2+}$ and/or Asc, however, no changes in sperm -SH-groups were detected when compared to controls. In another experiment, the activity of sperm binding to zona pellucida was evaluated through binding to salt-stored porcine oocytes. In control and Asc treatment groups, sperm binding to zona pellucida were significantly (P ＜ 0.05) higher than in medium with Fe$^{2+}$. On the other hand, there is not a significant increase in binding to zona pellucida with spermatozoa treated by Fe$^{2+}$/Asc. In summary, the present study suggests that Fe$^{2+}$/Asc causes an enhancement in fertilizing ability that is associated with penetration rate increased without change of spermatozoa binding capacity to homologous zona pellucida.o homologous zona pellucida.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.2
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• pp.69-75
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• 2021

Cryopreservation is a widely-used efficient means of long-term sperm preservation. However, unlike other types of semen, cryopreserved boar semen has reduced fertility and the efforts continue to optimize post-thawing sperm recovery. In this study, we evaluated the effects of various washing solutions (Hulsen solution, lab-made DPBS and commercial DPBS) on post-thawing porcine sperm kinematics (CASA system), viability (SYBR-14/PI) and acrosome integrity (PSA/FITC). We also examined the effect of washing-centrifugation on frozen-thawed semen kinematics. The results indicate that type of washing solution and post-thawing centrifugation alters parameters linked to sperm quality (total motility, progressive motility, viability and acrosome integrity). Significantly higher (p < 0.05) motility and progressive motility were obtained when cryopreserved semen was processed with Hulsen solution. The post-thaw percentage of live and intact acrosomal sperm was significantly higher in group 1 (Hulsen solution) as compared to other groups. Following thawing-centrifugation, the results showed significantly higher motility and progressive motility in group 1 than other groups. However, the latter two DPBS groups did not differ statistically. Taken together, Frozen-thawed spermatozoa motility, acrosome integrity and viability can be affected by the type of washing solution used. Moreover, centrifugation of frozen-thawed semen has an unfavorable effect on total motility and progressive motility.

• Journal of Life Science
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• v.26 no.8
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• pp.943-947
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• 2016

The objective of this study was to see if fairly simple magnetic nano-particle treatment enhances boar semen qualities. Boar semen samples were prepared from the swine AI center and samples were divided by 4 different motility groups (1, >90%; 2. 80~90%; 3. 70~80%; 4. <70%) using computer assisted sperm analysis (CASA) evaluation. Boar semen was extended using BTS extender and same number of magnetic nano-particles as total number of spermatozoa in each sample was treated for 20 min and collected for 5 min at room temperature. Sperm qualities such as motility and viability were evaluated by the CASA before and after treatment. Sperm abnormality and degree of agglutination were also evaluated under the microscopic examination before and after treatment. There were significant changes (p<0.05) on sperm motility from all 4 different groups in the average of 7.11% after treatment. The enhancement of sperm motility changes was more clear in the groups of lower sperm motile groups (<70% and 70~80%; 19.12±1.08% and 5.67±0.71%, p<0.05). The sperm motility character in terms of curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP) and linearity (LIN, %) showed also similar pattern but motility enhancement wear more clear in below 70% motile group. Average sperm viability was increased to 4% by magnetic nano-particles (p<0.05). The percentage of sperm abnormality was also reduced significantly (p<0.05) to the range of 3.7~4.5% before after treatment. The degree of sperm agglutination was also reduced in lower motility groups by the magnetic nano-particle purification.

• 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.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.9
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• pp.251-258
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• 2020

This study investigated the effect of Zardaverine supplementation in freezing extender, on kinetic characteristics of post-thawed boar sperm. Cryopreservation of boar sperm is an important technique of assisted reproductive technology and genetic resource banking. Although this technique is particularly useful, freeze-thaw cycles associated with sperm cryopreservation significantly reduce sperm quality. Semen from mature Duroc boars were collected and cryopreserved in freezing extenders (LEY) treated with varying concentrations of Zardaverine (0, 20, 50, 75, 100 𝜇M). The time-dependent kinetic characteristics of post-thawed spermatozoa were determined after thawing by applying computer-assisted sperm analysis (CASA). We observed that the motility immediately after thawing was significantly higher in 20 𝜇M stocks than in control (0 𝜇M) and the other treatments (p<0.05). Curvilinear velocity (VCL) in 0 𝜇M and 20 𝜇M stocks were significantly higher than the other treatment groups, except 75 𝜇M (p<0.05). Higher average path velocity (VAP) was obtained at 20 𝜇M as compared to 100 𝜇M, whereas amplitude of head lateral displacement (ALH) was significantly higher at 20 𝜇M than 50 𝜇M and 100 𝜇M (p<0.05). No differences were obtained for Straight-line velocity (VSL) and Linearity (LIN). In conclusion, our results indicate that Zardaverine improves the motility, VCL, VAP, and ALH of post-thawed boar sperm.

• Korean Journal of Animal Reproduction
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• v.24 no.3
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• pp.255-262
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• 2000

The aim of this work was to study the effects of ascorbic acid (Asc) and/or ferrous sulfate (Fe$^{2+}$) and spernatozoa preincubation on the in vitro fertilization in porcine. Porcine follicular oocytes matured in culture were inseminated with frozen-thawed boar spermatozoa preincubated for 0, 1, 2, 3, 4 and 5h. The penetration rates (37~51%) were not significantly different between durations of spermatozoa preincubation in medium with 0.1 mM Asc. The addition of 1.0 mM Fe$^{2+}$ during spermatozoa preincubation were not significantly affecting the penetration rates (41~56%). When spermatozoa were preincubated with Asc and Fe$^{2+}$, the penetration rates had a tendency to increase with time of spermatozoa preincubation, and were significantly (P<0.05) higher in spermatozoa preincubated with that than without Asc and Fe$^{2+}$ for 5 h. On the other hand, when spermatozoa were preincubated in fertilization medium without Asc and/or Fe$^{2+}$, the penetration rates were significantly (P<0.05) higher in medium with Fe$^{2+}$ than with Asc or Asc and Fe$^{2+}$ for in vitro fertilization. The rate of polyspermy in penetrated oocytes in medium with Asc and Fe$^{2+}$ decreased with the period of spermatozoa preincubation. Despite different culture conditions for spermatozoa preincubation, no differences were observed in polyspermy rates in the presence of Asc and/or Fe$^{2+}$ These results indicate the advantage of preincubating spermatozoa with Asc and Fe$^{2+}$ and an addition of Fe$^{2+}$ during in vitro fertilization with spermatozoa preincubated maintain penetration potential without increased polyspermy rates on in vitro fertilization in porcine oocytes.on in porcine oocytes.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2006.10a
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• pp.114-115
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• 2006

• Journal of Embryo Transfer
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• v.7 no.1
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• pp.41-47
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• 1992

1. In vitro system, LR and FSR accelerated and facilitated meiotic progression, and LH selectively improved cytoplasmic maturation which is required to promote the formation of a male pronucleus. 2. Caffeine (2mM) in the fetilization medium was required not only for inducing zona penetrating ability of boar also for developing to the male pronucleus of the penetrat- ing spermatozoa in vitro. 3. The germinal vesicle (GV)stage was observed for the first 17.6 hr;germinal vesicle break-down (GVBD)stage between 17.6~26.4 hr ;metaphase I (M-I)from 26.4 - 30. 9hr;anaphase I(A-I)ranged from 30. 9~33.4hr;telophase I(T-I) at 33.4~34.4hr; and metaphase II(M-II) at 34.4-48hr. 4. The addition of 10%(v /v) pig follicular fluid (pFF) to maturation media significantly increased the rate of nuclear maturation of pig oocytes (p<0.01), whereas the rate of nuclear maturation of pig oocytes among three different media did not differ. 5. The presence of a primary culture of POEC promotes in vitro development of early cleavage stage pig embryos.