Despite that porcine spermatogonial stem cells (pSSCs) have been regarded as a practical tool for preserving eternally genetic backgrounds derived from pigs with high performance in the economic traits or phenotypes of specific human diseases, there were no reports about precise definition of niche conditions promoting proliferation and maintenance of pSSCs. Accordingly, we tried to determine niche conditions supporting proliferation and maintenance of undifferentiated pSSCs for short-term. For these, undifferentiated pSSCs were progressively cultured in different composition of culture medium, seeding density of pSSCs, type of feeder cells and concentration of growth factors, and then total number of and alkaline phosphatase (AP) activity of pSSCs were investigated at post-6 day culture. As the results, the culture of 4x105 pSSCs on mitotically in activated 2x105 STO cells in the mouse embryonic stem cell culture medium (mESCCM) supplemented with 30 ng/ml glial cell line-derived neurotrophic factor (GDNF) was identified as the best niche condition supporting effectively the short-term maintenance of undifferentiated pSSCs. Moreover, the optimized short-term culture system will be a basis for developing long-term culture system of pSSCs in the following researches.

The current study was conducted to evaluate the biocompatibility of α-1,3 galactosyltransferase knockout pig bone graft in a rat calvarial defect model. Porcine cancellous bones were harvested from general and alpha-gal KO pigs and washed with 70% ethanol solution and normal saline. Bone pieces of the alpha-gal KO pig underwent a chemical treatment process to delipidize and deproteinize the bone. Bone graft particles were freeze-dried and stored at −70°C until use. Each bone graft was implanted into the rat calvarial defect in a fresh general pig, fresh transgenic pig, and chemical-treated pig bone group. There was no systemic adverse effect on hematology or necropsy findings in all groups at 1 week and 4 weeks. In the microcomputed tomography analysis, bone volume increased significantly in the chemical-treated transgenic pig bone group, whereas bone mineral density decreased significantly in the fresh general pig bone group compared with other groups. Histological evaluation showed cellular infiltration located at the margin of the bone graft particles, especially in the fresh general pig bone group. These results indicate that fresh general pig bone can elicit a greater local inflammatory response than fresh transgenic pig bone. Further, chemical-treated transgenic pig bone graft was less immunogenic than fresh bone graft. In conclusion, transgenic pig bone is a more biocompatible graft material. In addition, chemical treatment can reduce bone graft immunogenicity by delipidizing and deproteinizing bone.

비육돈사의 슬러리에 함유된 악취물질의 농도를 계절별로 비교하여 초지 및 농경지에 살포되는 분뇨의 악취강도를 예측하고자 수행되었다. 1.비육돈사의 슬러리에 함유된 휘발성유기물의 계절별 농도 비교페놀류 중 p-크레졸 농도는 봄, 여름 및 가을에 차이가 없었지만, 페놀, 인돌 및 스카톨 농도는 여름과 가을에 비해 봄에 높았다(p<0.05). 돼지 슬러리의 경우 p-크레졸과 스카톨이 악취강도에 큰 영향을 주기 때문에 계절에 따른 휘발성유기물의 악취강도 차이는 크지 않을 것으로 판단된다. 2.비육돈사의 슬러리에 함유된 휘발성지방산의 계절별 농도 비교휘발성지방산의 경우 악취강도가 높지 않은 단쇄지방산의 농도는 봄철이 여름과 가을보다 높았으나(p<0.05), 악취강도가 상대적으로 높은 이성체지방산의 농도는 계절별로 숫자적인 차이는 보였지만 통계적인 차이는 없었다(p> 0.05). 계절에 따라 휘발성지방산이 악취강도에 미치는 영향은 높지 않을 것으로 판단된다. 결과적으로 봄철에 악취물질의 농도가 더 높은 것은 온도, 환기 및 돈사내부로 유입되는 물의 양에 의한 것으로 추정되며, 여름철은 슬러리에서 생성되는 악취농도 보다 환기에 의해 휘산되는 양이 많아서 슬러리의 악취농도가 감소되거나 증가되지 않았을 가능성이 있는 것으로 판단된다. 이상의 연구결과를 종합하면 비육돈사의 슬러리를 초지 또는 농경지에 살포하였을 때 계절에 의한 악취강도의 차이는 크지 않을 것으로 기대된다.

The aim of this study was to develop the production of crossbred pigs suitable to the Korean consumers’demand. Crossbred (Jeju native pig × Landrace) pigs (n=175) at the age of 200 days, approximately classified into three slaughter weight groups (59~95 kg, 96~110 kg and 111~148 kg) were used to investigate the carcass traits and retail cuts characteristics. We have chosen longissimus dorsi, triceps brachii, biceps femoris, semimembranosus muscles, shoulder butt, thoracic vertebra (5-11th) and last thoracic vertebra for investigations of aforementioned parameters. The results showed that the 111~148 kg group had larger loin area, higher scores for meat color, quality and carcass grade than the other remaining groups (p<0.05). Weights of shoulder butt, rib, picnic shoulder, loin, belly, tenderloin, ham retail cuts, lean meat, fat and bone increased with increasing carcass weight (p<0.05). Ratios of shoulder butt, picnic shoulder, tenderloin, ham, lean meat, skin and bone in the 59~95 kg group were higher than that of the other groups (p<0.05). Ratios of belly in the 96~110 kg group were higher than that in the 59~95 kg group (p<0.05) but similar to the ratios of the 111~148 kg group. Total length, longest length and shortest length of belly increased with increasing carcass weight (p<0.05). Additionally, the longest width and shortest width of belly in 59~95 kg group were lower (p<0.05) than that in the other remaining groups. Length of shoulder butt increased with increasing carcass weight (p<0.05) whereas, the width of shoulder butt showed no differences between the groups. Weights of triceps brachii of picnic shoulder, biceps femoris and semimembranosus muscles of ham increased with increasing carcass weight (p<0.05). Overall, the crossbred pigs of 111~148 kg group possessed better quality and carcass grade however, the yield of belly in 96~110 kg group was higher than the other groups. Based on the consumption preference of Korean consumers therefore, the slaughter weight of crossbred pigs at ≥96 kg could produce higher economic benefit.

본 연구는 논토양에서 돈분액비의 시용이 화본과 및 두과 녹비작물의 생육과 양분공급량에 미치는 영향을 조사하기 위해, 화본과 녹비작물과 두과 녹비작물을 파종하여 돈분액비 시용유무에 따라 biomass 및 양분공급량을 조사하였다. 파종된 화본과 녹비작물은 호밀(Secale cereale L.)과 청보리(Hordeum vulgare L.)이었으며, 두과 녹비작물은 자운영(Astragalus sinicus L.)과 헤어리베치(Vicia villosa roth)이었다. 돈분액비를 투입함에 따라 녹비작물의 biomass는 유의성 있게 증가하였으며, 돈분액비 무시용구 대비 돈분액비 시용구의 biomass 증가율은 청보리(154%) > 호밀(138%) > 헤어리베치(45%) > 자운영(41%)순으로 높았다. 녹비작물의 양분공급량도 biomass와 마찬가지로 돈분액비를 투입함에 따라 유의성 있게 증가하는 경향이었으며, 돈분액비 무시용구 대비 돈분액비 시용구의 N, P2O5 및 K2O 공급량은 각각 205∼220, 109∼197 및 166∼184% 범위이었다. 돈분액비를 투입함에 따른 녹비작물의 biomass와 양분공급량 증가율은 전반적으로 두과 녹비작물보다 화본과 녹비작물에서 매우 우수한 경향이었다. 특히, 화본과 녹비작물의 증가된 양분공급량은 두과 녹비작물에 비해 약 3배 정도 더 높았다. 이상의 결과를 종합해 볼 때, 돈분액비 시용에 따른 양분이용효율은 화본과 녹비작물(호밀 및 청보리)이 두과 녹비작물(자운영 및 헤어리베치)에 비해 더 효과적인 것으로 판단된다.

The main purpose of this study is to estimate the effect of adding Tea-N-Tris (TES) to the freezing buffer for miniature pig sperm. In particular, we attempted to identify the association between the MMPs expression and the fertility and viability of frozen sperm from each extender (LEY (Lactose Egg-Yolk), TLE (TES + LEY), TFGE (TES + Fructose + Glucose Egg-Yolk)). In accordance with this, Hypoosmotic Swelling Test (HOST) respond test was the lowest among sperms frozen in LEY while the highest HOST respond was observed among sperms frozen in TLE. Furthermore, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of MMP-9 and MMP-2 was the highest in sperms frozen in LEY, Meanwhile, sperms from the TFGE and TLE group showed lower level of MMP-9 and MMP-2 expression in the order of TLE being the lowest. LEY group showed lower rate of blastocyst development than the TES supplement group, although the difference was not statistically significant. Meanwhile the rate of blastocyst development appeared similar when sperms from TLE and TFGE group were used for IVF. Together, these results indicate that adding Tea-N-Tris to the sperm freezing buffer only suppresses MMPs protein activation but also maximize in-vitro fertility, providing a means to improve the success rate in the in vitro manipulation of miniature pig sperm.

Low efficiency of somatic cell nuclear transfer (SCNT) is attributed to incomplete reprogramming of transfered nu-clei into oocytes. Trichostatin A (TSA), histone deacetylase inhibitor and 5-aza-2’deoxycytidine (5-aza-dC), DNA methy-lation inhibitor has been used to enhance nuclear reprogramming following SCNT. However, it was not known molec-ular mechanism by which TSA and 5-aza-dC improve preimplantation embryo and fetal development following SCNT. The present study investigates embryo viability and gene expression of cloned porcine preimplantation embryos in the presence and absence of TSA and 5-aza-dC as compared to embryos produced by parthenogenetic activation. Our results indicated that TSA treatment significantly improved development. However 5-aza-dC did not improve development. Presence of TSA and 5-aza-dC significantly improved total cell number, and also decreased the apoptot-ic and autophagic index. Three apoptotic-related genes, Bak, Bcl-xL, and Caspase 3 (Casp3), and three autophagic-re-lated genes, ATG6, ATG8, and lysosomal-associated membrane protein 2 (LAMP2), were measured by real time RT-PCR. TSA and 5-aza-dC treatment resulted in high expression of anti-apoptotic gene Bcl-xL and low pro-apoptotic gene Bak expression compared to untreated NT embryos or parthenotes. Furthermore, LC3 protein expression was lower in NT-TSA and NT-5-aza-dC embryos than those of NT and parthenotes. In addition, TSA and 5-aza-dC treated embryos displayed a global acetylated histone H3 at lysine 9 and methylated DNA H3 at lysine 9 profile similar to the parthenogenetic blastocysts. Finally, we determined that several DNA methyltransferase genes Dnmt1, Dnmt3a and Dnmt3b. NT blastocysts showed higher levels Dnmt1 than those of the TSA and 5-aza-dC blastocysts. Dnmt3a is lower in 5-aza-dC than NT, NTTSA and parthenotes. However, Dnmt3b is higher in 5-aza-dC than NT and NTTSA. These results suggest that TSA and 5-aza-dC positively regulates nuclear reprogramming which result in modulation of apoptosis and autophagy related gene expression and then reduce apoptosis and autophagy. In addition, TSA and 5-aza-dC affects the acetylated and methylated status of the H3K9.

Cathepsin B is abundantly expressed peptidase of the papain family in the lysosomes, and closely related to the cell degradation system such as apoptosis, necrosis and autophagy. Abnormal degradation of organelles often occurs due to release of cathepsin B into the cytoplasm. Many studies have been reported that relationship between cathepsin B and intracellular mechanisms in various cell types, but porcine embryos has not yet been reported. Therefore, this study evaluated the effect of cathepsin B inhibitor (E-64) on preimplantation developmental competence and quality of porcine embryos focusing on apoptosis and oxidative stress. The expression of cathepsin B mRNA in porcine em-bryos was gradually decreased in inverse proportion to E-64 concentration by using real-time RT-PCR. When putative zygotes were cultured with E-64 for 24 h, the rates of early cleavage and blastocyst development were decreased by increasing E-64 concentration. However, the rate of blastocyst development in 5 μM treated group was similar to the control. On the other hand, both the index of apoptotic and reactive oxygen species (ROS) of blastocysts were sig-nificantly decreased in the 5 μM E-64 treated group compared with control. We also examined the mRNA expression levels of apoptosis related genes in the blastocysts derived from 5 μM E-64 treated and non-treated groups. Expre-ssion of the pro-apoptotic Bax gene was shown to be decreased in the E-64 treated blastocyst group, whereas expre-ssion of the anti-apoptotic Bcl-xL gene was increased. Taken together, these results suggest that proper inhibition of cathepsin B at early development stage embryos improves the quality of blastocysts, which may be related to not only the apoptosis reduction but also the oxidative stress reduction in porcine embryos.

양돈 사료에 발효탄수화물을 첨가하여 돼지의 슬러리에 서 악취물질의 농도를 평가하였다. VFA 분석결과, 분뇨의 SCFA 농도는 대조구, 땅콩껍질, 아몬드피 및 골든화이버 구에서 각각 1,893, 1,591, 1,433 및 1,319 ppm으로 대조구 에서 가장 높았고(p<0.05), 아몬드피와 골든파이버 구에서 가장 낮았다(p<0.05). VFA의 구성을 살펴보면 SCFA 중에 서 아세트산의 비율이 가장 높으며, 다음으로 프로피온산, 부티르산, BCFA 순으로 낮았다. BCFA의 농도는 대조구, 땅콩껍질, 아몬드피 및 골든화이버 구에서 각각 98, 92, 78 및 74 ppm으로 대조구에서 가장 높았고(p<0.05), 골든화이 버 구에서 가장 낮았다(p<0.05). VOC 분석결과, 분뇨의 페 놀류 농도는 대조구, 땅콩껍질, 아몬드피 및 골든화이버 구 에서 각각 97.3, 47.0, 54.3 및 33.3 ppm으로 골든화이버 구 에서 가장 낮았으며, 대조구에서 가장 높았다(p<0.05). 그 리고 p-크레졸 농도가 페놀류 농도의 93~96%를 차지하였 다. 인돌류 농도는 대조구, 땅콩껍질, 아몬드피 및 골든화 이버 구에서 각각 1.8, 1.3, 1.2 및 1.0 ppm으로 발효탄수화 물 처리구간에 차이가 없었으며(p>0.05), 대조구에서 가장 높았다(p<0.05). NH4 +-N의 농도는 대조구, 땅콩껍질, 아몬 드피 및 골든화이버 구에서 각각 1,395, 995, 995 및 836 ppm으로 대조구에서 가장 높았으며, 골든화이버 구에서 가 장 낮았다(p<0.05). 가축의 장내 또는 슬러리에서 발효과정 동안 미생물이 성장을 위해 필요로 하는 에너지가 제한요 소로 작용되면, 미생물은 단백질을 에너지원으로 이용하기 때문에 많은 아미노산이 분해되어 악취물질이 증가 될 수 있다. 따라서 다른 처리구에 비하여 골든화이버 구에서 VOC의 농도가 가장 낮은 것은 골든화이버의 높은 NDF 함량으로(Getachew et al., 2004) 단백질 발효보다 탄수화 물 발효가 활발하게 일어나 악취물질의 농도가 낮았다고 판단된다.

본 연구는 양돈 슬러리의 악취물질 감소에 효과가 있는 것으로 확인된 물질의 효능을 비교분석하기 위하여 수행되 었다. 페놀류 농도는 고추냉이와 아몬드피 구에서 낮았고, 인돌류 농도는 아몬드피 구에서 가장 낮았다(p<0.05). 따라 서 슬러리에 아몬드피를 첨가하면 VOC 농도가 감소되었 다. SCFA 농도는 팽이버섯폐배지와 아몬드피 구에서 높았 고, BCFA 농도는 대조구에 비해 다른 모든 처리구에서 낮 았다(p<0.05). 또한 슬러리의 pH는 팽이버섯폐배지와 아몬 드피 구에서 낮았다(p<0.05). 따라서 팽이버섯폐배지 또는 아몬드피를 슬러리에 첨가하면 SCFA 농도는 증가되고 BCFA 농도는 감소되는 것으로 나타났다.

The aim of the present study was to evaluate the effect of ultimate pH of semimembranosus muscle on quality characteristics of dry-cured ham. The sample selection was determined based on ultimate pH of semimembranosus muscle, and samples were then classified into three groups: A (pH 5.61±0.09), B (pH 5.86±0.06) and C (pH 6.13±0.09). Our results depicted that the ultimate pH had a significant effect on the quality characteristics of dry-cured ham. Particularly, as the ultimate pH increased, the pH values of the dry-cured hams significantly (p<0.05) increased while weight loss decreased. Significantly (p<0.05) higher CIE a*, b* and chroma values were observed in the dry-cure hams from the samples with lower ultimate pH. Additionally, the values of some texture characteristics (e.g., hardness and chewiness) significantly (p<0.05) decreased as the ultimate pH increased. However, no significant differences among the three pH groups were observed for water activity, salinity, volatile basic nitrogen (VBN), calorie as well as sensory properties of dry-cure hams (p>0.05). These results clearly demonstrate that the ultimate pH of semimembranosus muscle is more related to quality characteristics than sensory attributes of dry-cured ham.

국산밀 재배시 가축분뇨 액비를 시용하여 생육에 미치는 영향과 사료가치를 분석하여 우리밀 생산물의 소비다양화 가능성을 알아보기 위하여 본 실험을 수행하였다. 용도별 우리밀을 선발하고 청보리로 사용되고 있는 영양보리를 대조구로 파종하여 생육 및 사료가치를 조사하였다. 각 작물별 파종 15일후 유묘출현율은 액비 처리구 화학비료 및 무처리구에 비해 2.4배 높았다. 또한 시험작물 모두 초장이 액비시용구에서 약 2배 이상 증가하였다. 월동 후 최고분얼기에는 밀이 영양보리에 비해 초장은 0.7∼1.8cm 작았으나 분얼수는 1.5∼2개가 많았다. 생체중의 경우 액비처리구에서 시험에 사용된 모든 밀이 영양보리에 비해 1.01∼1.11배 높았고 연백밀이 5,102kg 10a-1로 가장 많았다. 사료가치중 단백질 함량은 액비처리로 보리와 모든 밀 품종에서 유의성있게 증가하였고 조경밀이 9.19 %로 가장 높았다. NDF와 ADF 모두 청보리와 대등하거나 높았다. TDN은 금강밀이 69.86∼72.11 %로 가장 높았고 고소밀이 64.61∼66.07 %로 가장 낮았다. 시험전후 토양 특성은 가축분뇨 액비처리를 하였을 때 EC, 유기물, 유효인산 및 치환성 양이 함량이 증가하여 화학비료와 대등한 결과를 얻었다. 이상의 결과로 국산밀을 재배할 때 가축분뇨액비를 적절히 활용할 경우 재배단가를 낮추고 곡물밀의 공급이 과잉일 경우 사료용으로 공급하여 국산밀의 소비안정성에 도움을 줄 수 있을 것으로 판단된다.

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal α -1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from α-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% CO2 incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers (CD45－, 29+, 90+ and 105+) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited CD45－, 29+, 90+ and 105+ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at 1×103 cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs (15.0 ± 0.4 μm) had a little larger cell size than Wild BM-MSCs (13.5 ± 0.3 μm). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG 0.2 μM, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.

A total of 30 Korean native pigs (gilt 15, boar 15) were used to investigate the carcass characteristics, meat quality, amino acid, and fatty acid composition by gender. The carcass weight of boars were significantly higher than gilts, whereas the carcass yield of gilts had significantly higher than boars (p<0.01). Boars had significantly higher moisture contents in loin muscle than gilts, whereas the protein contents of loin muscle had significantly higher in gilts than boars (p<0.01). In the results of meat quality analysis, the cooking loss (p<0.01), shearing force (p<0.05), lightness (L) and yellowness (b) in meat color (p<0.05) were significantly higher, but the pH was significantly lower (p<0.01) in gilts compared with boars. Arginine (p<0.05), alanine, aspartic acid, histidine, leucine, lysine, phenylalanine, serine, threonine and tyrosin (p<0.01) for gilts were significantly higher than those for boars. The results of fatty acid composition showed that gilts had significantly higher contents of C16:1n7, C18:1n9, C20:1n9 (p<0.01) than boars in intramuscular fat, whereas boars had significantly higher contents of C18:2n6, C20:4n6 (p<0.01) and C18:3n3 (p<0.05) than gilts in intramuscular fat.

The objective of this study was to determine the effect of post-activation treatment with cytoskeletal regulators in combination with or without 6-dimethylaminopurine (DMAP) on embryonic development of pig oocytes after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT). PA and SCNT oocytes were produced by using in vitromatured pig oocytes and treated for 4 h after electric activation with 0.5 μM latrunculin A (LA), 10.4 μM cytochalasins B (CB), and 4.9 μM cytochalasins D (CD) together with none or 2 mM DMAP. Post-activation treatment of PA oocytes with LA, CB, and CD did not alter embryo cleavage (85.8～88.6%), blastocyst formation (30.7～ 32.4%), and mean cell number of blastocysts (33.5～33.8 cells/blastocyst). When PA oocytes were treated with LA, CB, and CD in combination with DMAP, blastocyst formation was significantly (P<0.05) improved by CB+DMAP (42.5%) compared to LA+DMAP (28.0%) and CD+DMAP (25.1%), but no significant differences were found in embryo cleavage (77.5～78.0%) and mean blastocyst cell number (33.6～35.0 cells) among the three groups. In SCNT, blastocyst formation was significantly (P<0.05) increased by post-activation treatment with LA+DMAP (32.9%) and CD+DMAP (35.0%) compared to CB+DMAP (22.0%) while embryo cleavage (85.5～85.7%) and blastocyst cell number (41.1～43.8 cells) were not influenced. All three treatments (LA, CB, and CD with DMAP) effectively inhibited pseudo-polar body extrusion in SCNT oocytes. The proportions of oocytes showing single pronucleus formation were 89.6%, 83.9%, and 93.3%, respectively with the increased tendency (P<0.1) by LA+DMAP and CD+ DMAP compared to CB+DMAP. Our results demonstrate that post-activation treatment with LA or CD in combination with DMAP improves pre-implantation development of SCNT embryos and the stimulating effect of cytoskeletal modifiers on embryonic development is differentially shown depending on the origin (PA or SCNT) of embryos in pigs.

본 연구는 농촌진흥청 국립축산과학원에서 사육중인 제주재래돼지와 랜드레이스의 상호교차교배를 통해 생산된 417두의 F2집단을 대상으로 성장형질(생시체중, 3주령 체중, 10주령 체중, 20주령 체중)을 측정하고, 개체별 혈액으로부터 Illumina Porcine SNP 60k BeadChip을 이용하여 유전자형 분석을 실시하여 성장형질과 유전체 전장의 단일염기다형의 유전자형 간에 연관성을 알아보았다. 단일연기다형의 유전자형 분석은 돼지의 성염색체를 제외한 18개의 상염색체 내에 나타난 52,574개의 SNP표지인자 중다형성이 나타나지 않은 7,564개의 표지인자, 모든 개체에서 이형으로 나타난 47개 표지인자 및 결측률이 10 % 이상인 1,830개의 표지인자가 나타나 분석에 앞서 사전 제거를 실시하였으며, 남은 44,133개의 표지인자를 체중형질과 표지인자간 연관성 분석하는데 이용하였다. 체중에 영향하는 고정효과에 대해 일반선형모형을 설정하여 사전 보정을 실시하였으며, 여기서 얻어진 잔차값을 이용하여 표지인자와 월령별 체중간의 연관성 분석을 실시하였다. 분석결과 전장의 유전체 정보 중 통계적 판단의 오류를 고려하여 연관성이 강한(p <10-6)표지인자가 생시(BWB), 3주령(BW3), 10주령(BW10) 및 20주령(BW20) 체중에서 각각 657개, 846개, 49개, 122개로 추정되었다. 생시와 3주령 체중에 공통으로 유의적 영향을 하는 표지인자가 286개로 나타났으며, BWB와 BW10에서 5개, BWB와 BW20에서 8개, BW3와 BW10에서 13개, BW3과 BW20에서 11개, BW10과 BW20에서 1개로 나타났다. 또한 염색체별 성장형질에 영향하는 표지인자의 분포를 조사한 결과, 주령별 체중에 영향하는 표지인자는 전장의 유전체에 고르게 분포하는 것으로 조사되었으며, 특히 생시 및 3주령 체중에 영향하는 표지인자는 특정 염색체(SSC9)에서 고도의 통계적 유의차(p <10-15)를 나타내는 유전자 좌위가 있는 것으로 추정되었다.

Artificial insemination (AI) has been performed widely in swine industry using fresh liquid sperm instead of frozen type of sperm. However fresh sperm are not able to preserve more than three days with optimal motility and other sperm parameters for the successful fertilization, since in vitro stored sperm has an oxidative stress that resulted increase of abnormality and acrosome reation. To overcome these major problems, novel preservative formulation is needed to neutralize the oxidative stress and to provide suitable physiological environment for sperm in in vitro. In this study, naturally derived substances such as Poncirus trifoliate (Trifoliate orange), Garcinia mangostana (Mangosteen), pig placenta and testis extracts were tested as sperm preservative agents. Placenta extracts (PE), trifoliate orange extracts (TOE), testes extracts (TE) and mangosteen extracts (ME) were applied to analyze specific parameters for sperm motion characteristics individually and combinatorial. Each individual extract treatment can accelerate the sperm motility but noticeably TOE, TE and ME treatments exhibited the considerable and significant preservation of sperm motility. PE, TE and ME showed a significant (p<0.05) increase in ALH after one week. Further we evaluated the five different combinations of these extracts on sperm motility and its motion characteristics. Surprisingly even after one week ME, TOE and TE combination significantly preserved the sperm motility about 75%. It is noteworthy that unlike individual extract treatment, combination of ME, TOE and TE simultaneously protect the sperm motility and its motion characteristics. Taken together these data conclude that addition of ME, TOE and TE can be effective for preservation of pig sperm.

In this study, we used flow a cytometric assay to evaluate plasma membrane integrity and mitochondrial activity in post-thawed sperm that was supplemented with ginsenoside-Rg1. Varying concentrations of ginsenoside-Rg1 (0, 25, 50 and 100 μM/ml) were used in the extender during cryopreservation to protect the DNA of thawed sperm, thereby increasing the viability and motility rate as evaluated using a computer-assisted sperm analysis (CASA) method. The results derived from CASA were used to compare the fresh, control, and ginsenoside-Rg1 groups. Sperm motility and the number of progressively motile sperm were significantly (p<0.05) higher in the 50 μM/ml ginsenoside-Rg1 group (61.0±4.65%) than in the control (46.6±7.02%), 25 μM/ml (46.2±4.76%), and 100 μM/ml ginsenoside-Rg1 (52.0± 1.90%) groups. However, the velocity distribution of post-thawed sperm did not differ significantly. Membrane integrity and MMP staining as revealed using flow cytometry were significantly (p<0.05) higher (91.6±0.82%) in the 50 μM/ml ginsenoside-Rg1 group than in the other groups. Here, we report that ginsenoside-Rg1 affects the motility and viability of boar spermatozoa. Moreover, ginsenoside- Rg1 can be used as a protective additive for the suppression of intracellular mitochondrial oxidative stress caused by cryopreservation.

일반적으로 돼지는 어떤 종류의 모색을 가지든 피부는 연분홍색을 띄는 것으로 알려져 있으나, 경 남 김해 H 육가공회사로 출하된 개체 중 흑모색에 검은색 피부를 가진 돼지가 발견되었다. 그 원인 을 규명하고자, 모색과 연관되어 있다고 알려진 MC1R, KIT 유전자와 피부색과 연관되어 있다고 알 려진 ASIP 유전자의 특징을 살펴보았다. MC1R의 sequencing 분석 결과, 아미노산 67, 68번째 자 리의 6개 염기 C(CCC CCC)는 Hampshire와 동일한 ED2/ED2 유전자형인 것으로 밝혀졌고, KIT의 경우 qOLA_CNV, Pyro_Splice 및 sequencing 분석한 결과, Duroc의 i/i 유전자형과 같은 유전자형 으로 밝혀졌다. ASIP의 경우 염기서열을 분석한 결과, 모든 종에서 차이가 없는 것으로 확인되었다. 유연관계 분석을 위해 Phase software와 network 분석을 실시한 결과, MC1R은 Hap22와 Hap23 이, KIT는 Hap22, Hap43과 유색종과 유연관계를 형성하는 것으로 확인되었다. 반면에 ASIP는 Hap04, Hap09이 야생멧돼지와 중국재래돼지를 제외한 품종들과의 유연관계를 확인할 수 있었다. 더 나아가 각 품종 간 유사성 분석을 위해 PCA를 실시한 결과, MC1R은 Berkshire, KIT는 Berkshire와 Hampshire가 유사성을 가지는 것으로 밝혀졌고, ASIP는 Berkshire 와 Duroc의 유사 성을 관찰할 수 있었다.