• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.123-126
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• 2008

The objective of this study was to determine the potential hazardous effects of sorting process by flowcytometry on the quality of boar spermatozoa by flowcytometer. Freshly collected boar semen was diluted and divided into two groups; control none sorted and sorted. Sperms in sorted group were processed with flowcytometer for cell sorting with $100\;{\mu}M$ nozzle under the 20 psi pressure. Measurements on each parameter were made at two time points, 0hr (right after sorting) and 24 hr post sorting. Although there was a tendency of lower viability in sorted group than none sorted control group, the percentage of live cells in control ($75.83{\pm}6.92\;&\;59.53{\pm}10.34$) was not significantly different from sorted ($59.70{\pm}7.37\;&\;43.97{\pm}3.76$) at both 0 and 24 hr post sorting. However, sorted sperm showed significantly lower mitochondrial function compared to the control at both 0 h ($79.37{\pm}3.22\;vs.\;63.50{\pm}10.05$) and 24 hr ($67.27{\pm}3.22$ vs. $46.97{\pm}5.37$) time points (p<0.007). Sperm DNA fragmentation rate was significantly lower in control ($22.0{\pm}7.04$) than that of sorted ($32.27{\pm}7.49$) at 24 hr time point (p<0.0002). Taken together, these data suggested thatsorting process by flowcytometer may have influenced sperm motility rather than viability. Also high speed sperm sorting by flowcytometer has significant effects on DNA fragmentation on elapsed time after sorting.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.219-224
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• 2006

The objective of this study was to investigate the toxic effects of Bisphenol A(BPA) and di-2 ethyhexyl phthalate (DEHP) as endocrine disrupters on sperm motility, viability, membrane integrity and abnormality during in vitro incubation of boar semen. Semen were randomly divide into 24 groups and healed with different concentrations of BPA md DEHP($1{\sim}100{\mu}M$) for 3, 6 and 9 hrs, respectively. The percentages of sperm motility and viability decreased by treatment time with both BPA and DEHP, and obiously differ from the controls. The percentages of sperm motility and viability significantly decreased by incubation with both $100{\mu}M$ of BPA and DEHP compared to control and other treatment groups(p<0.05). Sperm membrane integrity was significantly reduced by incubation with 10 and $100{\mu}M$ of BPA and DEHP, respectively(p<0.05), but sperm abnormality were not significantly affect both BPA and DEHP. These results indicate that high concentration of BPA and DEHP($>10{\mu}M$ can affect noxiously the sperm characteristics.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.9
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• pp.1475-1481
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• 2019

Objective: The objective of the present study was to investigate pork quality from Hereford${\times}$Berkshire female and intact male pigs reared outdoors in an alternative production system. Methods: Berkshire purebred sows were artificially inseminated, once in the fall and again in the spring of the following year, with semen from Hereford boars and managed free of antibiotics in an outdoor hoop structure until the last month of pregnancy, after which they were moved to a pasture-based unit of 0.8 hectares with individual lots with a farrowing hut, shade, and water ad libitum. Piglets were weaned at 4 weeks of age and housed in a deep-bedded hoop structure, grouped by sex. Animals were harvested at market weight of 125 kg, approximately 200 days of age. Hot carcass weight was collected at the time of the harvest. After 24 hours of refrigeration, carcass characteristics were measured. Longissimus dorsi samples collected from the right side loin. Loins were cut into 2.54-cm thick chops and were used to measure marbling score, color score, drip loss, and ultimate pH. Sensory panel tests were conducted as well at North Carolina State University. For pork characteristics and sensory panel data, trial and sex were included in the statistical model as fixed effects. Hot carcass weight was included in the model as a covariate for backfat thickness. Results: Neither the subjective nor the objective color scores displayed any differences between the boars and the gilts. No difference was found for pH and marbling score between trials or sexes. Gilts had a thicker backfat measurement at the last lumbar and a narrower longissimus muscle area measurement when compared to the boars. The only difference in the sensory characteristics was found between the trials for texture and moisture scores. Conclusion: Consumers were not able to detect boar taint under the condition of this study, which is that the intact males were reared outdoors. Additional trials would be necessary; however, based on the results of the present study, outdoor rearing can be suggested as a solution to the issue of boar taint.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.9
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• pp.116-123
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• 2018

The sperm quality is determined by the kinetic characteristics and acrosome integrity of the sperm. In the previous studies, analysis of semen quality had large errors because those experiments by using microscope had been conducted by people. In recent years, the molecular biological methods have been newly developed to complement the previous techniques. The ZAR1 gene is known to be a gene that affects early embryonic development in vertebrates, but there is no study of the association with semen. In this study, we analyzed the association between the kinetic characteristics and ZAR1 single nucleotide polymorphism (SNP) genotype. To detect the SNPs, we performed sequencing using genomic DNA from the whole bloods of Duroc pigs. We identified an SNP in the ZAR1 gene g.2540T>C. ZAR1 SNP genotypeing in 105 pigs revealed that the major and minor alleles were T and C, respectively. After we analyzed the association between the kinetic characteristics of sperm and the ZAR1 SNP genotype, we found a significant association in MOT (p<0.01), VSL (p<0.05) of the kinetic characteristics in the Duroc boars. It was confirmed that the boars with T allele were lower in MOT and VSL than C allele. Therefore, pigs with C allele are judged to be better at the MOT and VSL of semen. Based on these results, ZAR1 SNP genotyping may be a useful molecular biomarker to improve semen quality by applying molecular breeding technology.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.215-223
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• 2003

This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

• Journal of Animal Science and Technology
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• v.49 no.6
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• pp.729-736
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• 2007

The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(－0.23~－0.92 and －0.68~－0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.

• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.29 no.4
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• pp.150-157
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• 2018

The Zygote arrest 1 (ZAR1) gene is known to affect early embryonic development in various vertebrates. In this study, we performed the association analysis to check whether there is any significant relationship between semen kinematic characteristics and the ZAR1 gene. To determine semen kinematic characteristics, we measured motility (MOT), straight-line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), straightness (STR), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF) of spermatozoa in boars. In order to detect single nucleotide polymorphisms (SNPs), we extracted genomic DNA from multiple Duroc boars, and then subsequently used them in sequencing reactions. As a result, three SNPs were detected in the intronic region of ZAR1 gene (g.2435T>C in intron 2, g.2605G>A and g.4633A>C in intron 3 ). SNPs g.2435T>C and g.2605G>A were significantly associated with MOT (p<0.01) and VSL (p<0.05), and g.4633A

• Journal of Embryo Transfer
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• v.27 no.2
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• pp.107-111
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• 2012

Miniature pig sperm cryopreservation is continually researched in biotechnology for breed conservation and reproduction. It is important to control the temperature at each stage of cryopreservation and cryoprotectant. It is also necessary to find the optimal cryoprotectant concentration and chemical elements of the extender. Recently, many studies have used various cryoprotectant materials, such as dimethyl sulphoxide (DMSO), ethylene glycol (EG), antifreeze protein (AFP), amides, and glycerol. Glycerol is a commonly used cryoprotectant. However, glycerol has critical cytotoxic properties, including osmotic pressure and it can cause irreversible damage to live cells. Therefore, We focused on membrane fluidity modifications can reduce cell damage from freezing and thawing procedures and evaluated on the positive effects of trehalose to the viability, chromatin integrity, and motility of boar sperm. Miniature pig sperm was separated from semen by washing with modified- Modena B (mMB) extender. After centrifugation, the pellet was diluted with the prepared first extender. This experiment was designed to compare the effects that sperm cryopreservation using two different extenders has on sperm chromatin. The control group used the glycerol only and it was compared with the glycerol and glycerol plus trehalose extender. Sperm viability and motility were evaluated using WST1 assays and computer-assisted semen assays (CASA). Chromatin structure was examined using acridine orange staining. For the motility descriptors, trehalose caused a significant (p<0.01) increase in total motility ($57.80{\pm}4.60%$ in glycerol vs. $75.50{\pm}6.14%$ in glycerol + trehalose) and progressive ($51.20{\pm}5.45%$ in glycerol vs. $70.74{\pm}8.06%$ in glycerol + trehalose). A significant (p<0.05) increase in VAP ($42.70{\pm}5.73{\mu}m/s$ vs. $59.65{\pm}9.47{\mu}m/s$), VSL ($23.06{\pm}3.27{\mu}m/s$ vs. $34.60{\pm}6.58{\mu}m/s$), VCL ($75.36{\pm}11.36{\mu}m/s$ vs. $99.55{\pm}12.91{\mu}m/s$), STR ($54.4{\pm}2.19%$ vs. $58.0{\pm}1.63%$), and LIN ($32.2{\pm}2.05%$ vs. $36.0{\pm}2.45%$) were also detected, respectively. The sperm DNA fragmentation index was 48.8% to glycerol only and 30.6% to glycerol plus trehalose. Trehalose added group showed higher percentages of sperm motility, stability of chromatin structure than glycerol only. In this study, we suggest that trehalose is effective in reducing freezing damage to miniature pig sperm and can reduce chromatin damage during cryopreservation.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.4
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• pp.489-493
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• 2003

Reproductive traits of sows are determined chiefly by the genetic background of the dam herself. Whether the breed of the serviced boars also contributes is not yet clear. A total of 8,946 litters of Landrace (L) sows from 68 farms were examined. Of these, 876, 1,260, and 6,810 litters were produced from natural mating with L, Yorkshire (Y), and Duroc (D) breeds, respectively. The birth litter size (BLS), weaning litter size (WLS), live born piglets (LBP), and survival rate % (SR), of every litter were recorded. A general linear-model procedure was followed to estimate the effects of boars' breed (B), parity (P) and B${\times}$P interaction on the reproductive traits of the sows. Results show that BLS, WLS, and LBP were all significantly (p<0.001) affected by B and P. SR% was significantly influenced by B but not by P. L sows crossbred with Y or purebred with L produced litters with higher BLS, WLS, and LBP values than those bred with D. Pure L breeding yielded litters with lower SR% than did crossbreeding with D, while the difference between SR% due to LY and that due to LD crossbreeding was not significant. The interaction of B with P was significant with respect to BLS, WLS, and LBP (p<0.001), but not SR%. No significant B effect on reproductive traits was measured in sows at their first parity; but at latter parities, LL or LY produced litters with similarly high BLS, WLS and LBP, which values were all significantly greater than those of LD litters. The breed of boar evidently affected the subsequent reproductive performance of L sows and this effect may be further manipulated by the parity effect. Breed differences in semen quality and the success of fetus development with different interactions of the genetic background with the uterus function of the sow that may contribute to these effects are discussed.

• 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.