• Title/Summary/Keyword: 돼지정액

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Effects of Fertilization Time and Culture Medium of Pig Oocytes Matured In Vitro by liquid Boar Sperm Stored at $4^{\circ}C$ (체외성숙된 돼지난포란을 $4^{\circ}C$ 보존 액상정액으로 체외수정시 수정시간과 배양배지의 영향)

  • Park, C. S.;Y. J. Yi;Kim, M. Y.;Y. J. Chang;Lee, S. H.;D. I. Jin
    • 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.

Association with Kinetic Characteristics of sperm in Duroc Boar and the Zygote Arrest 1 gene Polymorphism (g.2540T>C) (Zygote arrest 1 유전자 변이(g.2540T>C)와 두록 정액의 운동학적 특성과의 연관성 분석)

  • Lee, Mi Jin;Ko, Jun Ho;Cho, Kyu Ho;Choi, Tae Jeong;Kim, Yong Min;Kim, Young Sin;Jin, Dong Il;Cho, Eun seok;Kim, Nam Hyung
    • 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.

Establishment of PCR Conditions for the Identification of Stenotrophomonas maltophilia Isolated from Boar Semen and Antimicrobial Susceptibility Patterns of the Isolates (돼지 정액에서 분리된 Stenotrophomonas maltophilia 확인을 위한 PCR 기법 개발 및 분리 균주의 항생제 감수성 양상)

  • Jung, Byeong-Yeal;Park, Bum-Soo;Kim, Ha-Young;Byun, Jae-Won;Kim, Ae-Ran;Jeon, Albert Byung-Yun;Kim, In-Cheul;Chung, Ki-Hwa
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1114-1119
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    • 2012
  • Bacteria are frequently contaminated during the collection and processing procedures of boar semen. Of the contaminants, Stenotrophomonas (S.) maltophilia is a Gram-negative bacterium that is widely distributed in a variety of habitats. Although PCR assays have been developed for the detection of S. maltophilia, they cross-react with some species of Xanthomonas. In this study, we designed a primer set for the detection of S. maltophilia in order to target the chiA (GenBank accession no. NC_010943) gene. The specific PCR products were amplified from S. maltophilia only, not from other tested strains that are frequently found in semen. The detection limit of the PCR was $1.5{\times}10^3$ CFU/ml with pure-cultured S. maltophilia and $1.5{\times}10^4$ CFU/ml with S. maltophilia spiked in semen. Twenty-six (5.9%) S. maltophilia were isolated from 440 semen samples. The PCR results exhibited 98.9% agreement with a comparison of S. maltophilia isolation. Also, the sensitivity and specificity of the PCR were 100% and 98.7%, respectively. In the antimicrobial susceptibility test, S. maltophilia isolates were highly susceptible to enrofloxacin and florfenicol, while the majority of them were resistant to amoxicillin/clavulanic acid, apramycin, ceftiofur, penicillin, and spectinomycin. These results indicated that the PCR using the chiA gene was proven to be reliable and effective for the detection of S. maltophilia with high levels of sensitivity and specificity.

The Reduction of Hydrogen Peroxide in Viable Boar Sperm Cryopreserved in the Presence of Catalase (Catalase 첨가에 따른 돼지 정액 동결 및 융해 후 생존 정자에서 Hydrogen Peroxide의 감소)

  • Kim, Su-Hee;Lee, Young-Jun;Kang, Tae-Woon;Kim, Yong-Jun
    • Journal of Veterinary Clinics
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    • v.28 no.1
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    • pp.13-19
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    • 2011
  • Semen cryopreservation induces the formation of reactive oxygen species (ROS), and the ROS cause sperm damage. We aimed to investigate the effects of the antioxidative enzyme catalase (CAT) on sperm quality and ROS during cryopreservation. Sperm rich fractions collected from five Duroc boars were cryopreserved in freezing extender with (200 or 400 U/mL) or without CAT (control). After thawing, sperm motility, viability, normal morphology, plasma membrane integrity, mitochondrial function and intracellular ROS were evaluated. CAT significantly improved total sperm motility at a concentration of 400 U/mL (P < 0.05), but didn't improve progressive sperm motility, viability, morphological defects, plasma membrane integrity and mitochondrial function in frozen-thawed boar sperm. In evaluation of ROS, CAT had no effect on reduction in ${\cdot}O_2$, but scavenged $H_2O_2$ in viable frozen-thawed boar sperm at concentrations of 200 and 400 U/mL (P < 0.05). In conclusion, CAT was not enough to improve quality of frozen-thawed sperm, but can reduce $H_2O_2$ generation in viable boar sperm during cryopreservation.

Toxicologic Effects of BPA and DEHP on Semen Characteristics in Pig (BPA와 DEHP가 돼지 정액 성상에 미치는 독성 영향)

  • Park, Dong-Heon;Jang, Hyun-Yong;Cheong, Hee-Tae;Kim, Jong-Taek;Park, Choon-Keun;Kim, Choung-Ik;Yang, Boo-Keun
    • 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.

Effect of Medium and Cumulus Cell on In Vitro Fertilization of Porcine Follicular Oocytes (배양액 및 난구세포가 돼지난포란의 체외수정에 미치는 영향)

  • Park, B.K.;Han, M.H.;Seo, K.W.;Park, C.S.;Lee, K.S.
    • Korean Journal of Agricultural Science
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    • v.23 no.2
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    • pp.206-211
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    • 1996
  • This study was conducted to investigate the effects of medium and cumulus cell on in vitro fertilization of porcine follicular oocytes. The results obtained are as follows ; 1. The normal fertilization rates of in vitro matured follicular oocytes cultured in 00, mT ALP and TCM-HEPES medium were 14.0~24.3%, 30.8~32.7% and 21.4~23.9%, respectively. These data indicated that the optimal medium for fertilization of porcine oocytes in vitro was the mTALP medium 2. The normal fertilization rates of epididymal sperm were 24.3%(80), 30.8%(mTALP) and 23.9%(TCM-HEPES), and those of ejaculated sperm were 14.0%(B0), 32.7%(mTALP) and 21.4%(TCM-HEPES). 3. The sperm penetration rates of cumulus-enclosed and cumulus-free oocytes on in vitro fertilization were 54.0% and 72.0%. The normal fertilization rates of cumulus-enclosed and cumulus-free oocytes were 11.9% and 21.5%. The normal fertilization rate of cumulus-enclosed oocytes was significantly(P<0.05) higher than that of cumulus-free oocytes.

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Studies on the In Vitro Fertilization and In Vitro Development of Porcine Embryos in Different Culture System (여러 가지 배양조건에서 돼지 난포란의 체외수정 및 체외발달에 관한 연구)

  • Kim, Jae-Young;Park, Hyang;Kim, Jae-Myung;Lee, Jung-Hyung;Park, Heum-Dae
    • Journal of Embryo Transfer
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    • v.19 no.1
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    • pp.19-25
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    • 2004
  • The objective of this study was to optimize the selection of sperm, optimal culture system of in vitro derived porcine embryos. The results obtained were summarized as follows: 1. When oocytes were inseminated with liquid sperm and frozen-thaw sperm, the cleavaged rate of liquid sperm (46.2%) was higher than that of frozen-thaw sperm (39.7%), however there were not show significant different each other. The blastocyst rates of liquid sperm (15.8%) was significantly higher than that of frozen-thawed sperm (9.3%)(P< 0.05). 2. When oocytes were inseminated with epididymal sperm after 1, 2 and 3 day storage, the cleavaged rate of epididymal sperm after 1, 2 and 3 day storage was 60.5, 61.0 and 56.8% respectively. The morulae (17.4, 19.9 or 17.3%) and blastocyst (8.7, 15.4, 11.3%) rate of epididymal sperm after 1, 2 and 3 day storage was no significantly respectively(P< 0.05). 3. In vitro developed to cleavaged rate of G1.3/G2.3 media used for culture was significantly(P< 0.05) higher as 62.1% compared with the results using the media NCSU23(52.8), however in vitro developed to blastocyst rate of NCSU23(11.6%) media was significantly(P< 0.05) higher than that'of G1.3/G2.3(4.7%). 4. When the fertilized oocytes were cultured with NCSU23 in addition to 1 mM glutathione(GSH), the cleavaged rate of treated groups of GSH(62.3%) was significantly higher than that of control(53.5%) respectively(P< 0.05). And in vitro developed to blastocyst rates of treated groups of GSH(15.6%) was higher than that of control(12.6%) however, there was no significant difference(P< 0.05).

질 협착증(strictures) 애견 인공수정(Artificial Insemination)으로 출산

  • 김영도
    • Journal of the korean veterinary medical association
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    • v.35 no.2
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    • pp.108-109
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    • 1999
  • 오늘날 `인공수정`하면 소,돼지는 일반화된 방법이 된 지 오래다. 그런데 `개의 인공수정`하면 아직도 많은 수의사나 번식가들이 회의를 가지고 있다. 필자 역시 KCRC 울산 지회로 가입하기 전까지는 그런 부류에 속했다. 그러나 KCRC의 수차례의 Canine Reproduction(AL)에 대한 세미나와 실습을 통하여 냉동 정액을 이용하여 인공수정을 할 수 있구나 하는 생각을 가지게 되었고 필자가 인공수정의 전 과정을 스르로 해 본 결과 나름대로 확신을 가질 수 있게 되었다. 물론 다음의 한 예가 인공수정에 대한 의문을 모두 제거하리라는 기대는 무리가 있겠지만 최소한 개의 인공 수정에 대한 이해의 폭은 넓히리라 확신한다.

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Utilization and Process of Frozen Semen in Boar (돼지 동결정액의 제조와 이용)

  • 임경순
    • Korean Journal of Animal Reproduction
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    • v.7 no.2
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    • pp.27-40
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    • 1983
  • Frozen boar semen should be utilized in swine production in order to get its advantages. Much studies were carried out for the practical use of frozen semen. Some of frozen boar semen are used in swine production and some companies are exporting frozen boar semen in U.S.A.. For the practical utilization of frozen semen in swine production in Korea, we have to get deep knowledge and understanding about frozen boar semen and studies on processing and conception of frozen semen.

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Studies on Semen Quality of Boar (돼지의 정액성상에 관한 연구)

  • 문승주;임계택
    • 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.

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