• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.119-124
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• 2002

This study was carried out to investigate the effects of packing materials of frozen boar semen to improve reproductive performance efficiency in pig. Boars were raised at Swine Artificial Insemination Center in National Livestock Research Institute, Sunghwan, Chungnam, Korea. We compared packing protocols for frozen boar semen among 5$m\ell$ maxi-straw, 5$m\ell$ cryogenic-vial, and aluminum-pack. Cryogenic-vial packing material showed similar sperm characteristics compared with maxi-straw packing material when the sperm was frozen above 15cm from liquid nitrogen and thawed at 52$^{\circ}C$ for 190 seconds. We investigated different thawing times to find out the optimal condition of freezing and thawing protocol with cryogenic-vial. Freezing above 15cm from liquid nitrogen and thawing at 52$^{\circ}C$ for 190 seconds were the optimal protocol compared with 120 and 150 seconds. However, normal acrosome rates did not show any differences among thawing times. Post-thawing results of maxi-straw in water at 52$^{\circ}C$ for 45 seconds had better total motility and curve linear velocity than those of cryogenic-vial in water 52$^{\circ}C$ for 190 seconds. However, there were no differences on straightness and normal apical ridge of sperm between maxi-straw and cryogenic vial. Non-return rate, farrowing rate and litter size of sows inseminated with frozen boar semen of commercial farms were higher in the maxi-straw than cryogenic-vial, but there were no significant differences between maxi-straw and cryogenic-vial. In conclusion, there were no significant differences between maxi-straw and cryogenic-vial and so, we may replace cryogenic-vial packing method instead of maxi-straw packing method by improvement of freezing and thawing rate.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.6
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• pp.793-798
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• 2006

The quality characteristics of semen are important indicators of the fertility of a boar. Development of genetic markers for the semen quality in boars will be beneficial to the improvement of porcine fertility. We investigated the relationship between the polymorphisms of epidermal growth factor (EGF), prostaglandin-endoperoxide synthase 2 (PTGS2) and prolactin receptor (PRLR) genes, and semen quality traits in boars. The genomic DNA of 233 boars (157 Duroc and 86 Landrace) from a central testing station was subjected to genotyping for surveying gene frequency. The EGF, PTGS2 and PRLR genotypes were determined using the restriction fragment length polymorphism method. Thirty-seven normal, mature Duroc boars from an AI center were also genotyped and their semen quality traits were collected. The effect of genotype on semen quality traits was analyzed by the least-squares means method using data corrected for season. The frequencies of the AA genotype of EGF, PTGS2 and PRLR in Duroc boars were 0.14, 0.01 and 0.66, respectively. In Landrace, the frequencies of the AA genotype were 0.03, 0.09 and 0.62, respectively. Boars with the BB genotype in EGF, with the AB genotype in PTGS2 and with the AA genotype in PRLR had significantly better semen quality with a higher percentage of normal sperm and a lower percentage of immature sperm than those with other genotypes. These findings imply that polymorphisms of EGF, PTGS2 and PRLR genes might be used as markers for improving the semen quality of boars.

• Journal of Life Science
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• v.23 no.9
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• pp.1133-1139
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• 2013

During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was $50pg/{\mu}l$ of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The $MIC_{90}$ values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and $16{\mu}g/ml$, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.

• 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.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.227-232
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• 2011

This survey was conducted to investigate the current status of swine artificial insemination(AI) centers registered as 'semen processing business' in Korea. The survey responses were collected by direct visitation or telephone conversation for 5 months from May through September in 2008. The survey showed that sixty-four AI centers were enrolled in local government and those of fifty-two AI centers were under operation. Forty-nine AI centers surveyed owned a total of 3,334 boars and the Duroc breed accounted for the highest rate(73.1%) of all boar breeds. In type of ownership, agricultural management corporations was the highest(42.3%) and followed by private ownership(34.6%). Large-scale AI centers in terms of own over 151 boar were surveyed as 5.9% and most AI centers own less than 100 boars(86.5%). The average number of boars per AI center was 68. The amount of liquid semen provided by 52 AI centers were 1,791,000 doses and each AI center provides average of 39,000 does, which is represented for 90% consumption by sows in Korea.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.2
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• pp.113-120
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• 2022

Sperm cryopreservation is a fundamental process for the long-term conservation of livestock genetic resources. Yet, the packaging method has been shown, among other factors, to affect the frozen-thawed (FT) sperm quality. This study aimed to develop a new mini-straw for sperm cryopreservation. In addition, the kinematic patterns, viability, acrosome integrity, and mitochondrial membrane potential (MMP) of boar spermatozoa frozen in the developed 0.25 mL straw, 0.25 mL (minitube, Germany), or 0.5 mL (IMV technologies, France) straws were assessed. Post-thaw kinematic parameters were not different (experiment 1: total motility (33.89%, 32.42%), progressive motility (19.13%, 19.09%), curvilinear velocity (42.32, 42.86), and average path velocity (33.40, 33.62) for minitube and the developed straws, respectively. Further, the viability (38.56%, 34.03%), acrosome integrity (53.38%, 48.88%), MMP (42.32%, 36.71%) of spermatozoa frozen using both straw were not differ statistically (p > 0.05). In experiment two, the quality parameters for semen frozen in the developed straw were compared with the 0.5 mL IMV straw. The total motility (41.26%, 39.1%), progressive motility (24.62%, 23.25%), curvilinear velocity (46.44, 48.25), and average path velocity (37.98, 39.12), respectively, for IMV and the developed straw, did not differ statistically. Additionally, there was no significant difference in the viability (39.60%, 33.17%), acrosome integrity (46.23%, 43.23%), and MMP (39.66, 32.51) for IMV and the developed straw, respectively. These results validate the safety and efficiency of the developed straw and highlight its great potential for clinical application. Moreover, both 0.25 mL and 0.5 mL straws fit the present protocol for cryopreservation of boar spermatozoa.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.165-174
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• 1999

Boar semen can be frozen successfully. However, there is a large variability in the extent of damage boar semen samples experiences during cryopreservation. This experiment was undertaken to find out factors that affect a post-thaw viability of boar spermatozoa. For this purpose, cryodiluents(BF5, LEY, Soejima and M-Soejima), cryoprotectants(glycerol. ethylene glycol, and propylene glycol), pre-freezing method(dryice-pellet, dryice-straw and L$N_2$vapour-st-raw) and total time required for freezing(2. 5, and 7 h) were compared as a factors. To investigate quality of semen during freezing process, motility(%), normal apical ridges(%, NAR), and proportion of living sperm(%) by flow cytometic analysis were assessed after collection, cooled, pre-frozen and post-thawing. Post-thaw motility of semen diluted with M-Soejima was 52.0%, respectively. When heparin, caffeine or heparin＋caffeine was added to 2nd cryodiluent of M-Soejima during freezing process, the highest motility after thawing was shown at the addition of caffeine (2mM), with 61.7$\pm$2.9% of motility. M-Soejima with heparin or caffeine was significantly higher than that of controI(p＜0.05). The result using glycerol(Gly), ethylene glycol(EG), propylene glycol(PG), and their mixture (Gly＋EG and Gly＋PG) as cryoprotectants, the highest motility was shown at the mixture treatment with Gly plus PG. However, the highest proportion of live spermatozoa was shown at Gly＋EG, there was no significantly difference among treatments(p＞0.05). When semen was pre-frozen with three manners(dryice-pellet, dryice-straw, and L$N_2$ vapor-straw), motility(%) of post-thaw spermatozoa was the highest in the L$N_2$ vapor-straw pre-freezing method of M-Soejima cryodiluent with 57.5% of motility, For a simple, economical and timesaving approach to freezing boar semen, total time required for freezing were 2, 5, and 7 hours, post-thaw motility were 43.8, 45.0 and 38.8%, NAR were 19.5, 22.7 and 28.5%, and viability were 20.8, 19.9 and 22.1%, respectively. This data suggests that boar semen diluted with M-Soejima cryodiluent contained caffeine, using mixture of glycerol and propylene glycol or ethylene glycol as cryoprotectants, frozen with 2 hours, can be taken better motility, NAR, and proportion of live spermatozoa.

• Journal of Embryo Transfer
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• v.24 no.1
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• pp.29-32
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• 2009

Prostaglandin $F_2{\alpha}$ ($PGF_2{\alpha}$) can facilitate release of epinephrine from the adrenal gland. The objective was to extend these findings and determine the effects of $PGF_2{\alpha}$ on sexual activity and semen collection training in sexually inexperienced boars. Boars (n=32; $281{\pm}18$ days of age) were moved individually once weekly to a semen collection room equipped with an artificial sow. Before entering the semen collection room, boar received i.m. treatments of $PGF_2{\alpha}$ at doses of 5 (n=8), 10 (n=8), or 20 (n=8), and control boar (n=8) were not treated. Reaction time (elapsed time after entering collection pen until the start of mounting) for boars receiving 5mg ($3.3{\pm}0.9\;min$), 10mg ($3.3{\pm}0.8\;min$) $PGF_2{\alpha}$ was shorter (p<0.05) than for controls ($6.7{\pm}0.9min$). Duration of ejaculation (min) per session was longer (p<0.05) for $PGF_2{\alpha}$ (10 mg, 20 mg)-treated boars ($7.3{\pm}0.7\;min$, $6.9{\pm}0.7\;min$), compared to control ($3.4{\pm}0.8\;min$). The number of training session per boars was less (p=0.056) for $PGF_2{\alpha}$ 10mg-treated boars ($1.0{\pm}0.4$), compared to control ($2.0{\pm}0.4$). Semen characteristic such as volume, concentration, the number of total ejaculated sperm, were similar for $PGF_2{\alpha}$-treated and controls. There was no apparent difference on sperm movement characteristics (Mot: motility, VCL : curve linear velocity, VSL : straight line velocity, VAP : average path velocity, LIN : linearity) after semen preservation by collected with or without $PGF_2{\alpha}$ treatment. In summary, administration of $PGF_2{\alpha}$ in boars increased the sexual activity and facilitated the training boars to mount an artificial sow for semen collection, but did not affect semen characteristic.

• Journal of Animal Science and Technology
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• v.52 no.5
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• pp.407-412
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• 2010

The objective of this study was to determine the effects of dietary supplementation of whole garlic powder (WGP) on semen characteristics and blood antioxidant level in boars. For this study, nine Duroc boars of 12 months age were used. Semen and blood samples were collected for 13 weeks, once in each week. The boars were fed the basal diet (BD; control) or BD supplemented with 3% WGP. There were no significant differences in the semen volume and sperm concentration between control and WGP group on all collection weeks. However, total sperm number per ejaculate was higher in the WGP group than that in the control group on collection weeks 6, 7 and 8 (P<0.05). Also, on collection weeks 5, 6, 7 and 8, mean of total ejaculated sperm numbers per boar were significantly higher in the WGP group compared to control group (P<0.05). On the other hand, ejaculation frequency per boar (boar's libido) and total ejaculated sperm number per boar were significantly increased in the WGP group compared to the control group, respectively (P<0.05). Although there was no difference in polyphenol level in seminal plasma between two treatment groups, polyphenol level in blood serum was significantly higher in the WGP group on collection weeks 9, 12 and 13 (P<0.05). These results indicate that dietary supplementation of 3% WGP improves boar libido and semen productivity such as ejaculation frequency per boar, total sperm number per ejaculate, mean of total ejaculated sperm number per head, and elevate the blood level of antioxidant (polyphenol) in boar serum.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.199-205
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• 2009

This study was carried out to investigate the effective genetic resources preservation system using the frozen boar semen. The porcine oocytes were matured for 44 hours in NCSU-23 medium with or without 10% Porcine Follicle Fluid (PFF), 0.5 ${\mu}g/ml$ porcine FSH, 0.5 ${\mu}g/ml$ equine LH, 1.0 ${\mu}g/ml$ 17 $\beta$-estradiol ($E_2$) and 10 ng/ml Epidermal Growth Factor (EGF) under mineral oil at $38.5^{\circ}C$ in humidified atmosphere of 5% $CO_2$ in air. After 44 h of culture, the oocytes were inseminated with frozen-thawed semen and fresh semen prepared with mTBM medium for 6 h. Later, set of 50 presumptive zygotes were transferred into 4-well dish (500 ${\mu}l$) of IVC medium. for embryos freezing, slow-freezing and vitrification methods were used as a cryopreservation. Differences among treatments were analyzed using General Linear Model Procedure by SAS Package (version 6.12) differences were considered significant when p<0.05. Following IVF and IVC, the rates of cleavage and blastocysts formation were significantly higher (p<0.05) in hormone supplemented group than that of hormone-free group (25.7 vs, 12.1). The development rates to cleavage and blastocysts were significantly higher in PZM-5 group than NCSU-23 group (60.3%, 46.6% vs 27.4%, 11.1%). Further improvement was achieved when PZM-5 was supplemented with FBS. Cleavage rates was significantly higher in fresh semen source group than frozen semen (66.7% vs 43.7%). However in blastocysts rates was similar two groups. Post-thaw survival rates of embryos were 1.2% and 2.2% in slow-frezing and vitrification groups, respectively. The results of our study suggest that it is still possible to improve the culture conditions and boar semen cryopreservation for enhance reproductive technology and animal genetic resources conservation.