• 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.

• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.141-146
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• 1990

12 crossbreed boars received 4 rations containing varying levels of lysine and DL-methionine. The results obtained from this study are summarized as follows ; 1. Semen volume, total sperm number of the treatment B, C and D were increased significantly (p<0.05) as compared with the treatment A but abnormal sperm percent of treatment B and C was decreased significantly (p<0.05) as compared with A. Sperm number and sperm mortility were not different from treatments. 2. Amino acids contents of sperm plasma were not different from treatments.

• 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.293-297
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• 2016

For evaluating the boar semen quality, sperm motility is an important parameter because the movement of sperm indicates active metabolism, membrane integrity and fertilizing capacity. Phospholipase C zeta (PLCz) is important enzyme in spermatogenesis, but the effect has not been confirmed in pigs yet. Therefore, this study was aimed to analyze their association with sperm motility and kinematic characteristics. DNA samples from 124 Duroc pigs with records of sperm 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 subjected. A SNP in non-coding region of PLCz g.158 A > C was associated with MOT (p < 0.05), VCL (p < 0.01), LIN (p < 0.01) and ALH (p < 0.05) in Duroc population. Therefore, we suggest that the intron region of the porcine PLCz gene may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

• 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.

• Reproductive and Developmental Biology
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• v.40 no.4
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• pp.33-37
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• 2016

This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.275-279
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• 2009

The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

• Reproductive and Developmental Biology
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• v.40 no.1
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• pp.1-5
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• 2016

This study investigated the effects of L-carnitine (LC) and nicotinic acid (NA) on sperm viability during liquid storage at $18^{\circ}C$ in miniature pigs. $10{\mu}M$ LC and 30 mM NA, combined LC and NA (LN) were treated in fresh semen for 3, 7, and 10 days. In results, sperm survival increased in NA- and LN-treated semen on 7 and 10 days (p<0.05), mitochondrial integrity of live sperm increased in LN-treated semen on 7 days (p<0.05), but not NA-treated semen. In addition, we examined the acrosome reaction of sperm in miniature pigs. LC and NA did not influence on acrosome reaction of boar sperm. In conclusion, LC and NA effectively maintained the viability and quality of sperm during long-term storage in miniature pigs, suggesting that the combined LN may be useful for improving the semen extender for long-term liquid storage in pigs.

• Reproductive and Developmental Biology
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• v.39 no.1
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• pp.7-11
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• 2015

Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to $3{\times}10^7/mL$ and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7~10 days.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.257-263
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• 2011

BPA, a diphenyl compound containing groups, that make it structurally similar to synthetic estrogen and is considered as one of the major endocrine disruptors. Silymarin has extensively been used to prevent and/or alleviate some human disease, especially for the treatment of adverse liver conditions. It has an antioxidative efficacy and cancer preventive efficacy. Therefore, we examined the hypothesis that silymarin can inhibit BPA-induced toxicity in boar sperm duing in vitro storage. Sperm characteristics (motility, viability, membrane integrity and mitochondrion activity) in semen exposed to BPA (10~200 uM) were sharply lowered, while it increase in a dose and time dependent manner due to silymarin addition (50~200 uM) into semen extender in the presence of BPA (100 uM). All of the evaluated characteristics were gradually improved in the groups that were treated with silymarin (50~200 uM) in the presence of BPA (100 uM) in comparison to BPA 100 uM alone group, irrespective of incubation periods (3 and 6 h). These results demonstrate that silymarin can ameliorate the toxicity of BPA on boar sperm characteristics during in vitro storage, suggesting that silymarin indirectly act as an antioxidant.