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Studies on In Vitro Fertilization and Development of In Vitro Matured Porcine Follicular Oocytes I. Effect of Various Media and Co-culture with Porcine Cumulus Cellsor Mouse Fetal Fibroblast Cells on In Vitro Development of In Vitro Fertilized Oocytes (체외성숙 돼지난포란의 체외수정과 배발달에 관한 연구 II. 각종 배양액, 돼지난구세포 및 생쥐태아간세포와의 공동배양이 체외수정 돼지 난포란의 체외발달에 미치는 영향)

  • 정형민;엄상준;승경록;이상준;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.113-120
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    • 1993
  • To provide the optimal culture conditions for the developm,ent of in vit개 produced embryos, we have been investigated various culture media as well as co-cultrue systems using porcine cumulus cells or mouse fetal fibroblast cells. Porcine ovaries were brought to the laboratory from local slaughter house within 1 hour after slaughtering and cumulus oocytes complexes were recovered from antral follicles(3~5mm) with 23 gauge needle. To maturate follicular oocytes, cumulus oocytes complexes were washed three times with TCM-199 containing 25mM HEPES and incubated(39$^{\circ}C$, 5% CO2 in air) in various maturation media for 42 hrs. Ejaculated and liquid storaged boar spermatozoa capacitated with different sperm capacitation methods and media were rpepared for fertilizing of matured follicular oocytes in vitro. Fertilization was performed by adding 5~10${\mu}\ell$ fo capacitated spermatozoa containing 1~5$\times$105 sperm/ml to droplets. Eighteen to twenty-eight hours after sperm insemination, fertilized eggs were washed three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three different media, m-KRB, BECM and TCM-HEPES were 0~1.0%, showing extremely lower rates. Especially, most of embryos were observed to arrest the development beyond 4-cell stages. The rates of embryos developed to 2-, 4-, 8-, 16-, 32-cell and morula or blastocyst stage in co-culture with porcine cumulus cells and mouse fetal fibroblast cells were 61.1~67.0%, 59.0~58.0%, 42.5~43.1%, 28.4~30.2% and 20.4~21.0%, respectively. These development rates upto morula or blastocyst stages were significantly higher than those of the embryos cultured in the basic culture medium(P<0.01). These findings suggest that co-culture of in vitro fertilized eggs with porcine cumulus cells or mouse fetal fibroblast cells enhance the development of fertilized eggs to morula or blastocyst stage in vitro.

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Studies on In Vitro Fertilization and Development of In Vitro Matured Porcine Follicular Oocytes I. Effect of Media and Capacitation Procedure on In Vitro Fertilization (체외성숙 돼지난포란의 체외수정과 배발달에 관한 연구 I. 배양액, 수정능획득 방법이 체외성숙 난포란의 체외성숙에 미치는 영향)

  • 정형민;엄상준;승경록;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.103-111
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    • 1993
  • These experiments were undertaken to establish the optimal culture systems for in vitro maturation, fertilization and subsequently embryonic development of porcine immature follicular oocytes isolated from the ovary of slaughtered pigs. Porcine ovaries were brought to the laboratory from local slaughter house within 1 hour after slaughtering and cumulus oocytes complexes were recovered from antral follicles (3~5mm) with 23 gauge needle. To maturate follicular oocytes, cumulus oocytes complexes were washed three times with TCM-199 containing 25mM HEPES and incubated (39$^{\circ}C$, 5% CO2 in air) for 42hrs. Ejaculated and liquid storaged boar spermatozoa capacitated with different sperm capacitation methods and media were prepared forfertilizaing of matured follicular oocytes in vitro. Fertilization was performed by adding 5~10${mu}ell$ of capacitated spermatozoa containing 1~5$\times$105 sperm/ml to droplets. Eighteen to twenty-eight hours after sperm insemination, fertilized eggs were washed three times with culture media and transferred to the culture media. The fertilization rates of in vitro matured follicular oocytes cultured in B. O., TCM-HEPES, m-KRB, and TALP-II media were 61.3%, 83.0%, 88.9% and 89.2%, respectively. In addition, the polyspermy rates were 60.7%, 66.5%, 53.8%, and 43.9%, respectively. These data indicated that the highest of fertilization and the lowest of polyspermy rate was shown in TALP-II medium. Spermatozoa capacitated by caffeine, heparin, and percoll density gradient treatment in the 4 different media, the fertilization rates were 33.0~57.2%, 39.9~90.2%, and 52.6~92.8%, respectively, showing the lowest rate in caffeine treatment. The development rate of follicular oocytes, fertilized with the spermatozoa capacitated by caffeine, heparin, and percoll gradient in the TALP-II medium, upto 2 to 4-cell stages were 32.6%, 74.5% and 70.9%, respectively. Finally, fertilization rates of follicular oocytes cultured with follicular fluid containing medium from 10 to 100% were 61.2~94.1% and the rates (90~94%) with 10~20% follicular fluids were significantly higher than those (85.3%) of cultured in the media without follicular fluid. In addition, the rates of pronucleus formation were also higher in follicular fluid treated group (73.1~83.0%) than those (64.7%) of oocytes cultured without follicular fluid. The highest fertilization and pronucleus formation rates was found in oocytes cultured with 10% follicular fluid. These results suggest that the addition of heparin or percoll density gradient method is better capacitation method. Furthermore, the addition of porcine follicular fluid to the fertilization medium may improve the fertilization rates and formation of pronucleus.

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Dietary Conjugated Linoleic Acid Can Decrease Backfat in Pigs Housed under Commercial Conditions

  • Dunshea, F.R.;Ostrowska, E.;Luxford, B.;Smits, R.J.;Campbell, R.G.;D'ouza, D.N.;Mullan, B.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1011-1017
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    • 2002
  • Conjugated linoleic acids (CLA) have been shown to decrease body fat content of individually-housed pigs but little is known about the responses under commercial conditions. Two studies were conducted to evaluate the effect of CLA under commercial conditions using contemporary genotypes. The experimental designs were similar between the two sites. Briefly, the studies were 2${\times}$2 factorial designs with the respective factors being sex (boar and gilt) and supplemental dietary CLA (0 and 4 g/kg). The studies involved 16-20 pens of pigs with 4-5 pens of each sex${\times}$CLA group. The first study was conducted with 144 pigs in 16 pens consuming a pelleted feed for 6 weeks at Bunge Meat Industries, Corowa, NSW. In the second study, 160 pigs were obtained from a commercial source and put into 20 pens in simulated commercial conditions and fed a mash diet for 7 weeks at Medina Research Station, WA. In Study 2 some aspects of meat quality were also investigated. Data from Study 1 showed that, although CLA had no significant effect upon feed intake and daily gain, the small changes in both resulted in a reduction in (-0.10 g/g, p=0.10) feed conversion ratio (FCR). While there was no significant effect of CLA on ultrasonic backfat depths, there was a significant decrease in carcass P2 (-1.0 mm, p=0.014) and estimated carcass fat (-7 g/kg, p=0.049). In the study conducted at Medina CLA had no significant effect upon feed intake, feed:gain or most measures of back fat. The exception was that dietary CLA decreased the rate of accumulation of fat at the shoulder, particularly in gilts, resulting in a significantly lower amount of shoulder fat at slaughter (-1.3 mm, p=0.044). CLA tended to increase dressing percentage although this was not significant (+0.5%, p=0.14). Meat from CLA treated pigs tended to be darker (p=0.12) and had a higher ultimate pH (p=0.06). These data suggest that under commercial conditions dietary CLA can improve growth performance and decrease P2 in pigs of an improved genotype, particularly gilts.

Effects of Grubbing by Wild Boars on the Biological Activities of Forest Floor (멧돼지(Sus scrofa coreanus Heude)의 임상 교란이 토양의 생물학적 활성에 미치는 영향)

  • Cha, Sangsub;Lee, Sang-Hoon;Chae, Hee-Myung;Shim, Jae Kuk
    • Korean Journal of Environment and Ecology
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    • v.26 no.6
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    • pp.902-910
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    • 2012
  • This study has been carried out to determine the effects of disturbances from wild boar grubbing on the functions of ecosystem. The experiments was performed in Mt. Jumbong of Long-term Ecological Research Sites of the Ministry of Environment. We measured soil physical properties, soil respiration($CO_2$), microbial biomass C, and soil enzyme activities from both disturbed and control plots. The disturbance sites were divided into two parts, mounds and pits. Soil organic matter contents were highest value at the control plots and lowest at the pit plots, respectively at 20.22% and 15.52%. The soil bulk densities were highest at the pit plots. Soil microbial biomass C and $CO_2$ evolution were significantly higher at the control plots compared to the disturbed plots. The results were positively correlated with soil organic matter contents. The cellulase activity and invertase activity in the soil showed similar pattern as the microbial biomass C and $CO_2$ evolution results. The cellulase activity and invertase activity in the soil were positively correlated with soil microbial biomass C. Soil organic matter contents seemed to affect the soil enzyme activities. The nitrate reductase activities were highest at the pit plots, which showed positive correlation with soil bulk density. The study results showed that the grubbing disturbances by wild boars induced the changes in soil properties, which affected soil microbial activities.

Studies on the Liquid and Freezing storage of Boar Semen III. Effects of Dilutor on Livability and Fertility of Liquid Semen and Effects of Dilution and Vessel on Livability of Frozen Semen (돈정액의 액상 및 동결보존에 관한 연구. III. 보존액이 액상정액의 정자생존성과 수태율에 미치는 영향과 희석방법과 용기가 동결정액의 생존성에 미치는 영향)

  • 임경순;정장용
    • Korean Journal of Animal Reproduction
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    • v.3 no.1
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    • pp.30-35
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    • 1979
  • A, B and C dilutors were used to make Ka (A plus B (1 : 1)) and Na (B plus C(1 : 1)) dilutors in this experiment. Three aliqots of semen were respectivly diluted 1 : 1 and 1 : 2 (semen: dilutor) with Ka, Na and C dilutors and stored at 5$^{\circ}C$ for 7 days in order to study their livability during storage. Fertility was checked for the diluted semen with Ka, Na and C dilutors. Whole semen and extended semen with Na dilutos with and without DMSO were cold shocked at various temperatures for 10 min. Effects of different 1st and 2nd dilution with A, B, C and Na dilutors and of vessels on freezability of spermatozoa were investigtigated. 1. Extended semen 1 : 2 with Na and C dilutors showed highest live sperm index during storage for 7 days at 5$^{\circ}C$. 2. The components of Na dilutor per 100$m\ell$ were skim milk 2.5g, trisaminomethane 0.54g, citric acid 0.265g, glucose 2.835g, fructose 1.5g, sodium lauryl sulfate, 0.08g, penicillin 0.06g, streptomycin 0.075g, and egg yolk 10$m\ell$. 3. Fertility of diluted semen was higher than that of whole semen. Ka dilutor showed higher fertility than Na and C dilutors, and there was no difference in the fertility between Na and C dilutors. 4. Na dilutor with DMSO showed slightly higher livability than Na dilutor without DMSO during storage for 7 days at 5$^{\circ}C$. 5. Cold shock at 1$0^{\circ}C$ for 10 min. decreased greatly the sperm livalility of whole semen but not of extended semen with Na dilutor. Addition of DMSO to Na dilutor has no effect in prevention of cold shock. 6. The extended semen with C. C dilutor (1st and 2nd dilution with C and C dilutor) showed higher post-thawing sperm livability than A.A and Na. B dilutors. Na. B dilution shwed higher post-thawing sperm livability than A.A dilution. There was no difference in the post-thawing livability between semen in 1$m\ell$ straw and 10$m\ell$ aluminium package.

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Crop Loss Survey by Wildlife in National Parks of Korea (한국 국립공원 내 야생동물과 농작물 피해)

  • Yoon, Seong-Il
    • Korean Journal of Environmental Biology
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    • v.25 no.3
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    • pp.223-227
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    • 2007
  • Crop loss and damage by wildlife are becoming more serious to residents inside and around National Parks of Korea annually. It could be caused by small size of Parks, which is couples of ten $km^2$ only, and more than a third of total parks' areas are belonging to privates. Wild boar (Sus scrofa) is one of the most harmful pests in nationwide. Treading down crop field using as bedding or resting site are major damaged types and foraging behavior is following. Besides of boars, Korean water deer (Hydropotes inermis), racoon dog (Nyctereutes procyonoides), and badger (Meles meles) are another serious pests. Mainly, farm products planted in upland, such as corns, sweet potatoes, cabbages, etc. are harmed easily, a rate reaches up to 76.9% of whole losses. 92.4% (n=187) of interviewees, who are mostly seniors, cultivate small crop fields smaller than 1 ha. Crop damage would be started from June to late November, time differences are happened due to latitude, climate condition, and crop differences of National Parks. Dusting animal repellents are easier and widespread methods to prevent animal foraging to crop fields and installing noisy makers using gun powder and setting up scarecrows are traditional methods for repelling animals. Nevertheless, effects of these self-defence are turned out to be useful soon after labors done only. Since legal compensation for crop damages by wildlife are not prepared systematically so far, crop loss are rarely reported to local government by most farmers. Systematic and objective methods for quantifying of crop loss by wildlife and population managements system should be offered urgently to well-managed animals habitat and residents' living inside National Parks.

Influence of the Substrate and Inhibitors Related to Phosphatidylinositol Metabolism in the Maturation Processes of Porcine Oocytes (돼지 난모세포의 성숙과정에서 Phosphatidylinositol 대사의 기질 및 억제인자의 영향)

  • 강승률;양보석;조인철;이성수;정진관
    • Journal of Embryo Transfer
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    • v.16 no.2
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    • pp.91-98
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    • 2001
  • We evaluated the effects of the substrate and inhibitors related to phosphatidylinositol metabolism on in vitro maturation and fertilization of porcine oocytes. Cumulus-oocyte complexes were cultured in mTLP-PVA medium supplemented with or without inositol (250 mM) fur 46h. Subsequently, these oocytes were inseminated with fresh boar semen in mTALP-PVA medium for 6h. At 6h after insemination, oocytes were cultured for further 12 h in TCM-199 supplemented with 10% FBS (fetal bovine serum). The higher percentage of oocytes in inositol-supplemented medium reached metaphase of the second meiotic division compared to those in control (81.4% vs. 67.3%; P<0.()5). following 18 h of insemination, more number of male pronuclei were formed in the oocytes matured in inositol-supplemented medium than in those of control experiment (42.0% vs. 27.3%; P<0.05). When oocytes were cultured in medium with 10mM LiCl (chloride lithium) or 0.5mM dbcAMP (dibutyryl cyclic adenosine monophosphate) to determine the role of inositol on the maturation of oocytes, these two drugs inhibited the meiotic division of oocytes (P<0.05). However, addition of inositol to the culture medium did overcome the inhibitory effect of these drugs on the oocyte maturation. DbcAMP and verapamil supplemented synergistically arrested the meiotic division of oocytes. Addition of verapamil did not inhibit germinal vesicle breakdown, but it severly inhibited the second meiotic division of oocytes. These results suggest that inositol exert its improving effects on maturation, by activating the PI (phosphatidylinositol) cycle and causing beneficial changes in both cytoplasm and membrane of oocytes.

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Ticks Collected from Wild and Domestic Animals and Natural Habitats in the Republic of Korea

  • Kim, Baek-Jun;Kim, Hyewon;Won, Sohyun;Kim, Heung-Chul;Chong, Sung-Tae;Klein, Terry A.;Kim, Ki-Gyoung;Seo, Hong-Yul;Chae, Joon-Seok
    • Parasites, Hosts and Diseases
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    • v.52 no.3
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    • pp.281-285
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    • 2014
  • Ticks were collected from 35 animals from 5 provinces and 3 metropolitan cities during 2012. Ticks also were collected by tick drag from 4 sites in Gyeonggi-do (2) and Jeollabuk-do (2) Provinces. A total of 612 ticks belonging to 6 species and 3 genera were collected from mammals and a bird (n=573) and by tick drag (n=39). Haemaphyalis longicornis (n=434) was the most commonly collected tick, followed by H. flava (158), Ixodes nipponensis (11), Amblyomma testudinarium (7), H. japonica (1), and H. formosensis (1). H. longicornis and H. flava were collected from all animal hosts examined. For animal hosts (n>1), the highest Tick Index (TI) was observed for domestic dogs (29.6), followed by Siberian roe deer (17.4), water deer (14.4), and raccoon dogs (1.3). A total of 402 H. longicornis (adults 86, 21.4%; nymphs 160, 39.8%; larvae 156, 38.9%) were collected from wild and domestic animals. A total of 158 H. flava (n=158) were collected from wild and domestic animals and 1 ring-necked pheasant, with a higher proportion of adults (103, 65.2%), while nymphs and larvae only accounted for 12.7% (20) and 22.2% (35), respectively. Only 7 A. testudinarium were collected from the wild boar (6 adults) and Eurasian badger (1 nymph), while only 5 I. nipponensis were collected from the water deer (4 adults) and a raccoon dog (1 adult). One adult female H. formosensis was first collected from vegetation by tick drag from Mara Island, Seogwipo-si, Jeju-do Province.

Identification of Quantitative Trait Loci(QTL) for Meat Color Trait on Chromosome 7 in Pig (돼지 7번 염색체에서 육색 연관 QTL 확인)

  • Choi, B.H.;Lee, H.Y.;Kim, T.H.;Hong, K.C.;Cheong, I.C.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.525-536
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    • 2004
  • The objective of this study was to identify the quantitative traits loci(QTL) for meat quality traits in pigs. Three-generation resource population was constructed from a cross between Korean native boars and Landrace sows. The resource population including founders, $F_1$ and $F_2$ was genotyped for 23 microsatellite markers on chromosome 7. The sex average total length of linkage map on chromosome 7 was estimated 154.6 cM. Meat quality traits including meat pH, meat color, drip loss, shear force, heating loss, crude fat, crude protein, crude ash and water content in muscle were collected from $F_2$ animals. For the QTL mapping, we used $F_2$ QTL Analysis Servlet of QTL express for web-based QTL mapping tools(http://qtl.cap.ed.ac.uk/). The QTLs for CIE-a and CIE-b on SSC7 were significantly detected at 1% and 5% chromosome-wide level, respectively.

Effects of Sperm Motility on In Vitro Production of Embryo and Correlation with Mitochondria Amount in Pig

  • Chung, Ki-Hwa;Kim, In-Cheul;Son, Jung-Ho
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.263-266
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    • 2010
  • Prediction of semen's fertilizing ability used in artificial insemination (AI) is one of very important factors on pig reproductive performance. In vitro fertilization (IVF) has been used for indirect evaluation of sperm's fertilizing ability and it has been showed as highly correlated index. In swine industry, increasing interest in preservation of boar semen raises questions on the sperm motility from semen used in commercial AI centers. Mitochondria in sperm mid-piece generate the energy to support motility and could be an explanation of impaired fertility. Objective of this study was to suggest usable sperm motility to farms in measuring the effect of sperm motility and sperm abnormality on in vitro production of embryo in which sperm's fertilizing ability can be determined indirectly. Semen samples were provided from local AI center and used within 3 days after collection. Semen samples were divided by 4 different motile groups (>70%; 61~70%; 51~60%; <50%) using CASA (computer-assisted sperm analysis) on the days of IVF. Developmental rate to the blastocyst stage from over 61% motile sperm group showed significantly higher rate than below 60% motile sperm group ($16.5{\pm}0.7{\sim}18.4{\pm}0.8%$ vs $6.3{\pm}0.8{\sim}11.5{\pm}0.7%$, p<0.05). In experiment to determine the relationship between sperm motility and viability and abnormality, over 61% motile sperm groups showed significantly higher viability rate compared to below 60% motile sperm groups ($84.8{\pm}4.0{\sim}88.1{\pm}4.0%$ vs $69.1{\pm}4.0{\sim}74.2{\pm}4.0%$, p<0.05). On the other hand, morphological sperm abnormality showed significantly higher in over 70% motile sperm group ($10.2{\pm}2.2$ vs $16.0{\pm}2.2{\sim}21.0{\pm}2.2%$, p<0.05). In experiment to find the correlation between sperm motility of 4 different motile groups and amount of mitochondria, lower motility group also showed lower level of mitochondria (p<0.05). The mitochondria parameter used in this study showed another possibility to differentiate the sperm motility. Taken together, because below 60% motile semen used in AI reduce the fertility, AI centers should provide the over 60% motile sperm to the farms at the time of AI.