• 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.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.84-84
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• 2002

The advantages of the OPS techniques(Vajta G et al, Mol Reprod Dev 51: 53-58,1998) give 1) high survival rates of various types of eggs, 2) quick and simple process, 3) inexpensive equipment and reduced chilling injury. The efficiency of IVM/IVF technique in the porcine species is relatively lower than that obtained in other species such as ruminants. Two experiments were designed to investigate the effects of in-vitro fertilization of porcine oocytes matures using different OPS protocol for chilling and warming of vitrification. Porcine oocytes from ovaries collected at abattoir were matured for 44 hours in TCM199 Earle's salt supplemental with pyruvate, pff, L-cysteine, hormones and gentamycin. Oocytes were denuded and fertilized with frozen boar semen by common method. Porcine embryos produced routinely by in-vitro culture system of NCSU23 medium. The vitrification and the warming were conducted by OPS method with the glass micropipette instead of straw vessels and modified the protocol of G.Vajta(1999). In Exp 1, Chilling/Warming:Holding Medium(HM)+EG+DMSO/HM +sucrose Medium(SM) at 39$^{\circ}C$ warm stage. In Exp 2, : PBS+CS+EG+Ficoll+ Trehalose/PBS+Trehalose at 25$^{\circ}C$ stage. Filling, freezing, packing, thawing out and further culturing were performed to follow the basic protocol of G Vajta. During IVM-lVC and post-warming, fertilization parameter and developmental potential were compared to and statistically analysed. It was not significantly different from Exp 1 and Exp 2 but 25$^{\circ}C$ of stage was slightly higher on the morula/blastocyst forming rate and better atmosphere for worker than that at 39$^{\circ}C$ stage.

• Journal of Embryo Transfer
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• v.21 no.4
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• pp.345-351
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• 2006

The objective of this study was to investigate the effects of preservation period of porcine liquid semen on bacterial contamination and in vitro production of embryo. Extended liquid semen was prepared by three mixture of boar's ejaculates from each farm without antibiotics, and were kept in $17^{\circ}C$ semen preservation incubator until use. Sperm motility was significantly (p<0.05) decreased as semen preservation time goes by (78.7$\pm$2.4% for 1 day vs. 71.1$\pm$2.4 and 64.8$\pm$2.4% for 3 and 5 days of presentation, respectively). Quantitative of bacteria in semen was significantly (p<0.05) higher in 5 days ($57.8\pm105.2\times10^4$ Cfu) compared to 0 and 3 days ($32.1\pm76.8\times10^4$ and $26.9\pm46.6\times10^4$ Cfu, respectively) of preservation. In terms of development of in vitro fertilization of porcine embryos inseminated by preserved semen, the rate of normal fertilization (2PN) was significantly (p<0.05) decreased in 5 days (56.0$\pm$2.6%) compared to 1 and 3 days (66.0$\pm$2.7 and 64.0$\pm$2.7%, respectively) of preservation. Cleavage rate was also significantly (p<0.05) affected by preservation period (75.0$\pm$4% for 1 day, 70.0$\pm$0.3 and 71.0$\pm$0.3% for 3 and 5 days, respectively). The in vitro developmental rate of blastocyst stage embryo was significantly (p<0.05) affected by semen preservation period (15.0$\pm$1.0% for 1 day vs. 11.0$\pm$0.9 and 8.0$\pm$0.9% for 3 and 5 days, respectively). It is concluded that more than 3 days of liquid semen preservation without antibiotics increased the quantity of bacteria resulted in detrimental effect on sperm motility and decreased both normal insemination rate and the developmental rate of blastocyst stage embryo.

• Journal of Embryo Transfer
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• v.30 no.2
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• pp.109-114
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• 2015

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.358A>T) of cluster-of-differentiation antigen 9 (CD9) gene reported to be significant association with MOT. Also, CD9 gene was expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as candidate gene for boar semen. This study was conducted to evaluate the pig SNP (g.358A>T) of CD9 gene as a positional controlling for semen parameters of post-thawed boar semen. To results, the g.358A>T SNP of the CD9 gene was significantly associated with the traits such as MOT, curve linear velocity, straight line velocity, average path velocity and amplitude of lateral head displacement. Particularly, the g.358A>T SNP significantly has the highest association with MOT and animals with AA genotype (p<0.001). Therefore, we suggest that the g.358A>T in the intron 6 region of the porcine CD9 may be used as a molecular marker for Duroc boar Post-thawed semen quality, although its functional effect was not defined yet.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.333-335
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• 1999

This study was carried out to find out the effect on fertilizing capacity according to sperm concentration of liquid boar semen. Four different doses with various motile sperm cells of 3.0$\times$10$^{9}$ , 2.5$\times$10$^{9}$ , 2.0$\times$10$^{9}$ , and $1.5\times$10$^{9}$ per 80$m\ell$ plastic bottle were inseminated twice 12 h interval after standing estrus in 6,818 sows. Farrowing rate and total piglets per litter were 82.2% and 10.9, respectively, with no significant differences among the other treatments. The presumption of optimal concentration of motile sperm cells in the liquid boar semen was best at 2.0~2.3$\times$10$^{9}$ per dose.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.23-30
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• 1990

These studies were carried out to investigate the effects of fetal calf serum(FCS), estrous porcine serum(EPS), porcine follicular fluid(PFF), hormone and matured cumulus cell(MCC) on in vitro maturation and fertilization of porcine follicular oocytes. The ovaries and testes were obtianed from slaughtered Landrace sow and boars, respectively. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3~5 mm and the semen were prepared from boar's epididymal cauda. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, EPS, PFF and MCC for 48hrs. in a incubator with 5% CO2 in air at 36$^{\circ}C$ and then matured oocytes were again cultured for 18~20 hrs. with $1.5\times$106/ml motile capacitated sperm in the modified Tyroide solution containing 100$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The maturation and fertilization rate of the follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS and PMSG＋HCG were 55.6~64.5% and 33.3~37.1%, respectively. 2. The maturation and fertilization rate of the follicular oocytes cultured in the TCM-199 medium supplemented with 20% EPS and PMSG＋HCG were 50.0~55.0% and 30.3~33.3%, respectively. 3. The maturation rate(59.0~64.2%) and fertilization rate(34.8~39.3%) of follicular oocytes cultured in TCM-199 medium supplemented 20% FCS and 50% PFF were higher than those of follicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and 10% and 50% PFF. 4. The maturation rate(60.0%) and fertilization rate(40.0%) of follicular oocytes cultured in TCM-199 medium supplemented with 20% FCS and granulosa cell (1$\times$106/ml) were significantly higher than those of fiollicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and granulosa cell.

• Journal of Embryo Transfer
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• v.17 no.1
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• pp.7-12
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• 2002

This study was carried out to investigate the possibility of porcine artificial insemination (A·I) on fertilizing capacity using intrauterine inseminator (IUI) method and conventional A·I (CAI) method. Number of sows used in this study was 15 far IUI and 59 fur (CAI), respectively. The results obtained are as fellows: 1 . The frozen and liquid semen used for A·I showed the higher farrowing rate in liquid semen (86.4%) than frozen semen (67%). Number of pigs born per semen type showed the higher values of number of piglets with no statistical significance using frozen semen (9.7) than liquid semen (9.3). 2. The farrowing rate per parity was highest in the 3∼5th parities (100%), f311owe4 by 0∼ 2th parities (60%), and was the smallest in 6 ∼ 10th parities (25%). Number of pigs born per litter was highest in 0∼2th parities (11.3), followed by 3 ∼ 5th parities (9.2) and lowest in 6∼ 10th parities. In the number of pigs bort per litter, the sow s in the high parities delivered lower number of piglets than those in low parities with no significant difference. These results indicated that fertilizing capacity could be improved by using IUI method.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.8
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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.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.287-291
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• 2016

For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Estrogen receptors 2(ESR2) is involved in estrogen related apoptosis in cell cycle spermatogenesis, but their functions have not been confirmed in pig until now. Therefore, this study was conducted to analyze their association with sperm motility and kinematic characteristics. DNA samples from 105 Duroc pigs with records of semen motility and kinematic characteristics [Total motile spermatozoa (MOT), Curvilinear velocity(VCL), Straight-line velocity(VSL), the ratio between VSL and VCL(LIN), Amplitude of Lateral Head displacement(ALH)] were analyzed. A SNP in coding region of ESR2 g.35547A > G in exon 5 was associated with MOT (p < 0.05) in Duroc population. Therefore, we suggest that the porcine ESR2 gene may be used as a molecular marker for Duroc boar semen quality, although its functional effects were not defined yet. These results might shed new light on the roles of ESR2 in spermatogenesis as candidate gene for boar fertility, but still the lack of association across populations should be considered.

• Korean Journal of Veterinary Research
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• v.45 no.3
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• pp.375-379
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• 2005

Porcine juvenile pustular psoriasiform dermatitis (PJPPD) is a disease of young pigs and characterized by nonpruritic round eruption of skin. The cause of this disease is yet undetermined but is presumed to be genetic predisposition. There may be few opportunities for veterinarian to detect this disease compared with actual situation in field because these lesions resolve spontaneously in two months. The authors detected spontaneous PJPPD case and performed clinical and pathological studies on three pigs from one farm. The specific skin lesions were observed in the forty-day old pigs of mixed breed, which were produced by the sows received semen from the same boar, restrictively. However, there was no skin lesion of pigs in suckling or fattening periods. Grossly, lesions were commonly found on the ventral abdominal part as a papule and were spreaded to the skin of whole body. With the spreading of lesions centrifugally, skin was showed as a umbilicated plaques or mosaic pattern with a few pustules or crusts. Microscopically, the most prominent lesion was the psoriasiform hyperplasia with acanthosis, down growth of rete ridges, exocytosis of eosinophils and neutrophils, ballooning degeneration of superficial epidermis, and koilocytic degeneration of keratinocytes. Additionally, there were moderate dermal edema and severe mixed cellular infiltration, especially eosinophils. No infectious agent which can cause the skin lesion, was detected or cultured, and no lesion caused by infectious agents was also observed, pathologically. With pathological results of this study, it is supposed that pathogenesis or severity of PJPPD may be related to the infiltration of eosinophil or hypersensitivity.