• 한국수정란이식학회지
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• 제31권2호
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• pp.117-121
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• 2016

Although piglets have been delivered by embryo transfer (ET) with in vitro produced (IVP) embryos and blastocysts, a success rate has still remained lower level. Unlike mouse, human, and bovine, it is difficult to a production of piglets by in vitro fertilization (IVF) because of an inappropriate in vitro culture (IVC) system in pig. Therefore, the present study was conducted to investigate whether minimized exposure time in IVC can improve the pregnancy and delivery rates of piglets. Immediately after IVM, the oocytes were denuded and co-incubated with freshly ejaculated boar semen for 3.5 to 4 hours at $38.5^{\circ}C$ under 5% $CO_2$ in air. To avoid long-term exposure to in vitro state, we emitted IVC step after IVF. After that the presumptive zygotes were transferred into both oviducts of the surrogate on the same day or 1 day after the onset of estrus. Pregnancy was diagnosed on day 28 after ET and then was checked regularly every month by ultrasound examination. The 3 out of 4 surrogates were determined as pregnant (75%) and a total of 5 piglets (2 females and 3 males) were delivered at $118.3{\pm}2.5$ days of pregnancy period. In conclusion, a short-term exposure time may be an important factor in the production of IVP-derived piglets. It can be apply to the in vitro production system of transgenic pig by IVF, cloning, and pronuclear microinjection methods.

• 한국가축번식학회지
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• 제19권2호
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• pp.147-152
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• 1995

본 연구는 돼지의 발정 동기화를 시키는데 있어 Altrenogest와 PG600간의 효과를 비교하기 위해 축산기술연구소에서 1994년부터 1995년 사이에 실시하였다. 먼저 시험돈은 자연 발정주기를 매일 2회씩 관찰하였고(Control), 한 group은 발정주기의 15일경에 PG600을 피하 주사했으며(PG600), 나머지 한 group은 발정주기와 관계없이 9일 동안 20mg의 Altrenogest를 사료에 첨가 급여하였다(Altrenogest). 그후 대조구를 제외한 두 그룹은 각각 PMSG 1500 IU를 PG600 처리후 16일째에, Altrenogest 처리구는 Altrenogest 처리후 24시간에 각각 근육주사하였으며, 약3일후 hCG 750 IU를 각 처리별로 근육주사하였다. 이 결과 대조구에서는 85%, Altrenogest는 90.1%, PG600구에서는 100%의 발정 발현율이 나타남으로써 전체 발현율로써는 처리간에 별차이가 없었으나, hCG주입후 9일내에 발정 발현정도는 대조구(25/53)에 비하여, PG600구 (47/47)가 높았다. 또한 황체수 및 회수난자수에서는 각각 대조구 12.9$\pm$1.8, 12.7$\pm$3.9, Altrenogest구 25.5$\pm$0.7, 15.0$\pm$4.2, PG600구 25.4$\pm$13.1, 19.0$\pm$12.8로 호르몬 처리간에는 별차이가 없다. 그러나 대조구에 비해서는 호르몬 처리구에서 유의적으로 (P<0.05) 높았으며, 정상 난자율에서는 호르몬 처리구보다 대조구에서 약간 높은 경향을 보였다. 따라서 본 연구에서 호르몬 처리에 의한 발정동기화가 효율적으로 가능하다는 것이 입증되었으며 이러한 발정동기화기술은 노동력 및 비용절감측면에서 효과가 있는 것으로 사료된다.

• 한국축산식품학회지
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• 제24권2호
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• pp.108-114
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• 2004

본 연구는 버크셔종 돼지의 출하체중과 성별에 따른 도체 및 육질 비교를 위해 실시하였다. 첨단양돈연구소에서 사육한 178-183일령 흑돼지(버어크셔) 72두를 공시하여 출하체중과 성별에 따라 95-104kg, 105-110kg 및 111-120kg으로 분리하여 조사하였다. 경남 김해시 어방동 소재 부경양돈 농협 도축장으로 수송하여 관행적인 방법으로 도축 후 도체 특성을 조사하였으며, 육질시험은 도축하여 24시간 냉각한 다음 등심부위(longissimus dorsi)를 공시하여 조사하였다. 출하체중이111-120kg 돼지가 95-104kg과 105-110kg 돼지보다 도체중과 등지방 두께는 증가하였고, 등급은 출하체중이 105-110kg과 111-120kg인 돼지가 A, B등급에 가까운 좋은 등급을 받았지만, 95-104kg인 돼지는 B, C 등급에다(p＜0.05). 돈육 p $H_{u}$, 육즙손실 및 가열감량은 출하체중과 성별에 따라 유사하였으며, 수퇘지가 암퇘지에 비해 전단력이 높았다.(p＜0.05). 돈육의 육색은 출하체중과 성별에 따라 유사한 경향이었다. 총 육색소 함량은 출하체중이 95-104kg인 암퇘지와 111-120kg인 수퇘지가 다른 출하체중과 성별보다 높았다. 조직특성은 출하체중에 따라 유사한 경향이었고, 미경산 암퇘지가 수퇘지에 비해 탄력성과 파쇄성이 높았다. 이상의 연구에서, 도체특성(도체중과 등지방두께)은 출하 무게와 성별에 따라 영향을 받았고, 암퇘지육은 수퇘지에 비해 전단력값과 조직특성이 향상되었다.다.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.185-191
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• 2010

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin and may participate in mammalian fertilization. Although correlations have been reported between reactive oxygen species (ROS) and sperm function, the relationship between PA activity and ROS is unknown. We determined the effects of ROS on sperm function and PA activities in boar spermatozoa preincubated under the X-XO system. When spermatozoa were treated with the X+XO group, a significant increase (p<0.05) was observed in the percentage of acrosome reacted spermatozoa compared with that of the control group. However, when antioxidants were added to the medium with X+XO, the rate of acrosome reaction tended to decrease. Also, a significantly lower percentage of acrosome reacted spermatozoa was observed in the X+XO+catalase group at 6 hr of incubation compared with that of X+XO group. The density of malondialdehyde (MDA) was higher in the X+XO group than in different treatment groups. In another experiment, incubation of spermatozoa in medium with X+XO was associated with a significant (p<0.05) increase in activity of tPA-PAI and tPA compared with the control group. Antioxidants decreased the increased activity of tPA-PAI and tPA by preincubation in the X-XO system. Also, a significantly lower (p<0.05) activities of tPA-PAI and tPA were observed in the X+XO+catalase group compared with the X+XO group. No significant differences, however, were observed in the activity of uPA. These results suggest that the increase of acrosome reaction by the X-XO system resulted in increase of PAs activity in the sperm incubation medium.

• Parasites, Hosts and Diseases
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• 제55권2호
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• pp.185-191
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• 2017

Tick is one of the most important arthropods in the transmission of vector-borne diseases. In this study, we investigated the abundance and species of ticks associated with swine and their habitats to assess the risk of spread of tick-borne diseases in host species, such as wild boars. Ticks were collected from 24 grazing or traditionally reared domestic pig farms and 8 habitats of wild boars in 8 provinces and 1 city in the Republic of Korea, by using the dragging and flagging methods. Ticks were also collected directly from 49 wild boars by using fine forceps. A total of 9,846 hard ticks were collected, including 4,977 Haemaphysalis longicornis, 4,313 Haemaphysalis flava, 508 Ixodes nipponensis, 1 Ixodes turdus, and 47 Amblyomma testudinarium. A total of 240 hard ticks were collected from 49 wild boars, including 109 H. flava, 84 H. longicornis, and 47 A. testudinarium. A total of 578 hard ticks were collected from areas around domestic pig farms. Only 2 hard tick species, 546 H. longicornis and 32 H. flava, were collected from these areas. A total of 9,028 hard ticks were collected from wild boars of 8 habitats, including 4,347 H. longicornis, 4,172 H. flava, 508 I. nipponensis, and 1 I. turdus. A. testudinarium was collected only from wild boars, and I. nipponensis and I. turdus were collected only from the habitats of wild boars.

• Parasites, Hosts and Diseases
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• 제55권2호
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• pp.207-212
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• 2017

Infections of Toxoplasma gondii and Babesia microti are reported in many wild animals worldwide, but information on their incidence and molecular detection in Korean wild fields is limited. In this study, the prevalence of T. gondii and B. microti infection in blood samples of 5 animal species (37 Chinese water deer, 23 raccoon dogs, 6 roe deer, 1 wild boar, and 3 Eurasian badgers) was examined during 2008-2009 in Gangwon-do (Province), the Republic of Korea (=Korea) by using serological and molecular tests. The overall seropositivity of T. gondii was 8.6% (6/70); 10.8% in Chinese water deer, 4.3% in raccoon dogs, and 16.7% in roe deer. PCR revealed only 1 case of T. gondii infection in Chinese water deer, and phylogenic analysis showed that the positive isolate was practically identical to the highly pathogenetic strain type I. In B. microti PCR, the positive rate was 5.7% (4/70), including 2 Chinese water deer and 2 Eurasian badgers. Phylogenetic analysis results of 18S rRNA and the ${\beta}$-tubulin gene showed that all positive isolates were US-type B. microti. To our knowledge, this is the first report of B. microti detected in Chinese water deer and Eurasian badger from Korea. These results indicate a potentially high prevalence of T. gondii and B. microti in wild animals of Gangwon-do, Korea. Furthermore, Chinese water deer might act as a reservoir for parasite infections of domestic animals.

• Journal of Animal Science and Technology
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• 제48권6호
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• pp.785-796
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• 2006

본 연구는 인공수정 센터에서 이용되고 있는 종모돈의 인공수정 분만율의 차이를 기준으로 하여 정액의 보존 일에 따른 정자의 생존율, CASA, HOST, IVP 및 SCSA를 조사하여 이들 정자의 분석법들과 인공수정 분만율과의 상관관계를 조사하여 종모돈의 수정 능력을 예측할 수 있는 정액 평가법을 개발하고자 실시하였다. 종모돈의 액상정액을 대상으로 CASA, HOST, IVP 및 SCSA 기법을 적용한 정액의 평가 결과는 다음과 같았다. 정액의 보존 일별 HOST 결과는 보존 일이 경과함에 따라 유의적으로 감소하였고(P<0.05), IVP 결과 난자 당 침투 정자 수는 보존 3일째에 유의적으로 감소하였다(P<0.05). 보존 0일과 6일에 HOST는 분만율이 80% 이상인 종모돈 군이 70% 미만인 종모돈 군 보다 유의적으로 높았고, 3일에서는 80% 이상인 종모돈 군이 다른 종모돈 군에 비하여 높았다(P<0.05). IVP는 보존 0일에 분만율이 80% 이상인 종모돈 군이 70% 미만인 종모돈 군보다 난자당 침투 정자 수가 유의적으로 많았다(P<0.05). %Red는 보존 0일과 3일에는 분만율이 80% 이상과 70% 미만인 종모돈 군 간에 유의적인 차이를 보였으나 보존 6일에서는 모든 종모돈 군간에 유의적인 차이가 있었다(P<0.05). %Red(r=0.79, P<0.01 ; r=0.86, P<0.01 ; r=0.88, P<0.01)는 정액의 보존 일이 증가함에 따라 분만율과의 부의 상관 계수가 증가하였다. 이상의 결과를 종합해 보면 SCSA는 정액의 수정 능력을 평가하는데 있어서 매우 유용한 방법이며, 정액의 보존 일은 여러 가지 정자 기능과 SCSA 결과에 크게 영향을 주는 것으로 나타났다. 정액의 수정 능력을 평가함에 있어서 정액의 보존 일령을 고려하고 정자 기능 검사와 SCSA를 병행하여 실시할 경우 종모돈의 수정 능력 평가 시 정확성을 더욱 높일 수 있는 유용한 방법이 될 수 있을 것으로 사료된다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.13-13
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• 2001

This study was undertaken to evaluate the effects of catalase using xanthine (X) - xanthine oxidase (XO) system on in vitro maturation and fertilization in pig. When follicular oocytes were cultured in maturation medium with X and/or XO, the maturation rates were not significantly different between in medium with and without catalase despite of different culture periods. However, significantly (P<0.05) higher maturation rates were obrained in culture with X-XO system. The rates of degenerated oocytes were increased with culture periods prolonged, and were significantly (P<0.05) higher in medium without than with catalase at 120 h of culture. On the other hand, the parthenogenetic oocytes were observed with high proportions at 72 h of culture, hut were not different in medium with and without catalase at various times of culture. In another experiment, the frozen-thawed boar spermatozoa treated with X-XO system for in vitro fertilization. The penetration rates were higher in medium with that than without catalase during the in vitro fertilization with, none (P<0.05), XO and X＋XO. On the other hand, when sperm were treated with none, X, XO and X＋XO, lipid peroxidation were higher in medium without that than with catalase. However, the changes in sperm penetration and lipid peroxidation showed opposite patterns. The sperm suspensions were also treated with X and/or XO for assay of sulfhydryl (-SH) group content. Under the above all conditions, sperm-SH group were higher detected In medium with that than without catalase. The activity of sperm binding to zona pellucida was also evaluated through binding to salt-stored porcine oocytes. In control group, sperm binding to zona pellucida were higher than in medium with X, XO and X＋XO groups. No significant differences, however, were observed between medium with and without catalase. In conclusion, the exposure of follicular oocytes and spermatozoa to X-XO system may be caused stimulating in vitro maturation and fertilization in pig. This work was supported by grant No. 2000-1-22200-001-3 from the Basic Research Program of the Korea Science & Engineering Foundation.

• 한국가축번식학회지
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• 제17권2호
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• pp.113-120
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• 1993

본 연구는 제외생산된 돼지 수정란의 처1외발생율을 제고하기 위하여 각종 배양액파 돼지난구세포 혹은 생 쥐태아간세포와의 공동배양 효과플 조사하였다 m-KRB, BECM 및 TCM-HEPES 배양액을 공시하 여 제외수정란을 배양한 결과 배반포기까지 발달하는 비율은 전처리구에서 0~1.0%로써 극히 저조하였다. 특히 대부분의 수정란은 4-세포기 단계에서 발달이 정지되었다. 한편, 단층세포가 유도된 돼지 난구세포나 생쥐 태아간세포와 함께 제외수정란을 공동배양한 결파 2, 4-, 8~16-, 32-세포기, 상실배가 빛 배반포로 받달하는 비율은 각각 61.1~67.0%, 59.0~58.0%, 42.5~43.1%, 28.4~30.2% 및 20.4~21.0%였다 이러한 결파는 단순배양액에서 체외배양한 수정란의 발탄 성적 보다유의하게 높은 것이었다. 이상의 결과를 종합하여 볼 때 1세포기 수정란을 체외에서 배양할때 체세포와의 공동배양은 수정란의 체외발달을 촉진하는 것으로 생각된다.

• 한국가축번식학회지
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• 제17권2호
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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.