본 연구는 솔잎 토착미생물을 이용하여 제조된 발효사료 급여가 돼지의 성장, 혈액성상, 도체형질 및 경제성에 미치는 영향을 규명하고자 실시하였다. 평균 체중 75kg인 3원교잡종 (Landrance× Yorkshire×Duroc) 180두를 공시하여 돈방 당 15두씩 시험구당 3반복으로 실시하였다. 발효사료는 원료 사료와 미생물을 혼합 후 혐기적 상태로 10일간 발효시켰으며, 양돈용 사료에 0 (대조구, CON), 3 (T1), 5 (T2) 및 10% (T3)로 대체하여, 42일간 급여 후 도축하였다. 발효사료의 조단백질 함량은 발효 후가 발 효 전보다 높았지만 (p<0.05), 조섬유 함량과 pH는 낮았다 (p<0.05). 돼지의 혈중 Hemoglobin 함량은 T3구가 대조구에 비해 높았고 (p<0.05), Hematocrit 함량은 대조구와 T1구가 T2구와 T3구에 비해 높았 고 (p<0.05), Platelet 함량은 T1구가 T2구와 T3구에 비해 높았다 (p<0.05). 대조구의 혈중 total cholesterol, LDL-cholesterol 및 Triglycerides 함량은 발효사료를 대체한 모든 시험구에 비해 높았으나 (p<0.05), HDL-cholesterol 함량은 적었다 (p<0.05). 지육중량과 지육율은 T1구와 T3구가 대조구와 T2 구에 비해 높았으며 (p<0.05), 등지방 두께는 T1구와 T2구가 T3구에서 비해 낮았다 (p<0.05). 육 등급은 발효사료를 대체한 모든 시험구가 대조구에 비해 개선되었으며 (p<0.05), kg 증체당 사료비는 발효사료 를 급여했을 때 약 2~6% 적었다. 따라서 솔잎 토착미생물을 이용하여 제조한 발효사료의 급여는 돼지의 건강과 육 등급 개선 및 사료비 절감에 도움이 될 것으로 사료된다.

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bcl-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MTT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG (2 μM) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

Two piglets and one juvenile pig were used to investigate closely what types of cells express green fluorescent protein (GFP) and if any, whether the GFP-tagged cells could be used for stem cell transplantation research as a middle-sized animal model in bone marrow cells of recloned GFP pigs. Bone marrow cells were recovered from the tibia, and further analyzed with various cell lineage markers to determine which cell lineage is concurrently expressing visible GFP in each individual animal. In the three animals, visible GFP were observed only in proportions of the plated cells immediately after collection, showing 41, 2 and 91% of bone marrow cells in clones #1, 2 and 3, respectively. The intensity of the visible GFP expression was variable even in an individual clone depending on cell sizes and types. The overall intensities of GFP expression were also different among the individual clones from very weak, weak to strong. Upon culture for 14 days in vitro (14DIV), some cell types showed intensive GFP expression throughout the cells; in particular, in cytoskeletons and the nucleus, on the other hand. Others are shown to be diffused GFP expression patterns only in the cytoplasm. Finally, characterization of stem cell lineage markers was carried out only in the clone #3 who showed intensive GFP expression. SSEA-1, SSEA-3, CD34, nestin and GFAP were expressed in proportions of the GFP expressing cells, but not all of them, suggesting that GFP expression occur in various cell lineages. These results indicate that targeted insertion of GFP gene should be pursued as in mouse approach to be useful for stem cell research. Furthermore, cell- or tissue-specific promoter should also be used if GFP pig is going to be meaningful for a model for stem cell transplantation.

The objective of this study was to evaluated the efficiency on sperm cryosurvival and ability of in vitro fertilization using Triladyl and Lactose Egg-Yolk(LEY) as extenders for cryopreservation of separated sperm by 65% percoll in miniature pig. Sperm viability was measured with SYBR-14/PI double stained sperm by flow cytometry. Ability on embryo cleavage rate and blastocyst development were observed by in vitro fertilization after frozen-thawing of sperm separated by 65% percoll. The experimental groups were designed that separated sperm by 65% percoll with Triladyl (ST) or LEY(SL) and unseparated sperm with Triladyl(UT) or LEY(UL) for cryopreservation. As a results, the viability was significantly(p<0.05) higher in ST(55.1%), SL(63.1%), UL(58.8%) than UT(38.2%) group. Sperm viability in SL(63.1%) group was significantly(p<0.05) higher than other experimental groups. On the other hand, embryo cleavage rate was significantly(p<0.05) higher in ST(79.1%), SL(83.2) than UT(74.1%) and UL(75.7%) groups at 96h after in vitro fertilization. Blastocyst development was also significantly(p<0.05) higher in ST(21.5%), SL(20.9%) than UT(17.0%) and UL (18.8%) groups. In conclusion, cryopreservation of miniature boar sperm separated by 65% percoll were beneficial to viability and capacity on in vitro fertilization.

The present study was conducted to investigate serotype distribution and biofilm formation of Actinobacillus (A.) pleuropneumoniae isolated from pneumonic lungs of pigs. A total of 37 A. pleuropneumoniae were isolated between January 2009 and June 2010. Serotypes of A. pleuropneumoniae isolates were determined using two different PCRs. The majority of isolates belonged to serotype 5 (n=31, 83.8%), and the others belonged to serotype 1 (n=4, 10.8%), and 2 (n=2, 5.4%), respectively. The ability of biofilm formation of the isolates was also determined by quantitative microtiter plate assay. Biofilm formation was observed in both 23 (62.2%) of the 37 field isolates and seven (43.8%) of the 16 reference strains. On the other hand, biofilm formation was various according to the serotypes: 20 (64.5%) of serotype 5, and three (75.0%) of serotype 1. However, two isolates of serotype 2 did not produce the biofilm in this study. Consequently, A. pleuropneumoniae serotype 5 was the most frequently detected (83.8% of the isolates), and 23 (62.2%) of 37 isolates exhibited biofilm positive phenotype in this study.

본 연구는 돼지의 육성기 또는 비육기에 Bio 이온수 급여에 따른 성장, 혈액분석 및 육질 특성 평가를 위하여 실시하였다. 시험구는 Bio 이온수 무 급여구 (대조구), 육성기 급여구 및 비육기 급여구로 3시험 구를 두었으며, 각 시험구 당 33두의 3원교잡종 (Landrace☓Yorkshire☓Duroc) 돼지를 배치하였으며, 총 99두를 이용하여 사양시험을 수행하였다. Bio 이온수 급여는 육성돈과 비육돈의 성장과 사료효율에 영 향을 미치지 않았지만 (P>0.05), 비육기 급여구에서 일당증체량과 A등급 출현율이 높게 나타났다. 대조구 에 비해 Bio 이온수를 급여한 처리구에서 혈액성상 분석 결과 적혈구와 백혈구의 수치가 증가하였다 (p<0.05). 일반성분, 육색, pH, 육즙감량, 가열감량 및 전단력에서 유의적인 차이는 나타나지 않았다. 지방 산 분석 결과 육성기 급여구에서 포화지방산/불포화지방산 비율이 낮게 나타났고, 불포화지방산의 함량 비 율이 가장 높게 나타났다. 또한 가열육 관능검사에서 향과 전체적인 기호도에서 높은 점수를 획득하였다.

For the screening of Brucella antibodies in pig, 2,140 pig serum samples were collected from six slaughter house in Korea between 2006 and 2007. The Rose Bengal test (RBT) and serum agglutination test (SAT) were used for initial screening for specific antibodies to Brucella, and competitive enzyme-linked immunosorbent assay (C-ELISA) was used for confirmation of presence of serum antibody for Brucella. Overall, 575 (26.9%) samples resulted in seropositive in RBT. In SAT, 50 (2.3%) and 10 (0.5%) samples showed suspicious positive and positive reaction, respectively, however, all sera tested in this study showed a negative reaction in C-ELISA. SAT and C-ELISA might be applicable as a tool for screening of swine brucellosis.

The first morphogenetic event of preimplantation development, compaction, was required efficient production of porcine embryos in vitro. Compaction of the porcine embryo, which takes place at post 4-cell stage, is dependent upon the adhesion molecule E-cadherin. The E-cadherin through -catenin contributes to stable cell-cell adhesion. Rho-associated kinase (ROCK) signaling was found to support the integrity of E-cadherin based cell contacts. In this study, we traced the effects of ROCK-1 on early embryonic development and structural integrity of blastocysts in pigs. Then, in order to gain new insights into the process of compaction, we also examined whether ROCK-1 signaling is involved in the regulation of the compaction mediated by E-cadherin of cellular adhesion molecules. As a result, real-time RT-PCR analysis showed that the expression of ROCK-1 mRNA was presented throughout porcine preimplantation stages, but not expressed as consistent levels. Thus, we investigated the blastocyst formation of porcine embryos treated with LPA and Y27632. Blastocysts formation and their qualities in LPA treated group increased significantly compared to those in the Y27632-treated group (p < 0.05). Then, to determine whether ROCK-1 associates embryonic compaction, we explored the effect of activator and/or inhibitor of ROCK-1 on compaction of embryos in pigs. The rate of compacted morula in LPA treated group was increased compared to that in the Y27632-treated group (39.7 vs 12.0%). Furthermore, we investigated the localization and expression pattern of E-cadherin at 4-cell stage porcine embryos in both LPA- and Y27632-treated groups by immunocytochemical analysis and Western blot analysis. The expression of E-cadherin was increased in LPA-treated group compared to that in the Y27632-treated group. The localization of E-cadherin in LPA-treated group was enriched in part of blastomere contacts compared to that Y27632-treated group. ROCK-1 as a crucial mediator of embryo compaction may plays an important role in regulating compaction through E-cadherin of the cell adhesion during the porcine preimplantation embryo. We concluded that ROCK-1 gene may affect the developmental potential of porcine blastocysts through regulating embryonic compaction.

Decorin (DCN) is a member of small leucine‐grich proteoglycans which are ubiquitous components of the extracellular matrix. It regulates many physiological processes, such as matrix formation, collagen fibrillogenesis, angiogenesis, cancer growth, and cardiovascular diseases. It has been shown that DCN is expressed in the uterus during pregnancy and modulates implantation and decidualization for the establishment and maintenance of pregnancy in mice and humans. Expression of DCN in the uterine endometrium during pregnancy has not been investigated in pigs. Thus, this study investigated expression of DCN in the uterine endometrium during the estrous cycle and pregnancy in pigs. Uterine endometrial tissues were from day (D) 12 and 15 of the estrous cycle and D12, D15, D30, D60, D90, and D114 of pregnancy. Northern blot and real‐gtime RT‐gPCR analyses showed that expression of DCN mRNA was detected throughout the estrous cycle and pregnancy with the highest levels during mid pregnancy. In situ hybridization analysis showed that DCN mRNA was localized to both luminal and glandular epithelia during the estrous cycle and pregnancy and also to chorionic membrane during mid pregnancy in pigs. To determine whether endometrial expression of DCN was affected by the somatic cell nuclear transfer (SCNT) procedure, DCN mRNA levels in the uterine endometrium from gilts with SCNT embryos on D30 of pregnancy were compared with those from gilts with normal embryos using real‐gtime RT‐gPCR analysis. The result showed that DCN mRNA levels in the uterine endometrium were not significantly different between gilts with normal embryos and SCNT embryos. These results suggest that DCN may play an important role for endometrial tissue remodeling during mid pregnancy, and DCN expression is not affected by the SCNT procedure at the early stage of pregnancy in pigs.

본 연구는 축산물품질평가원에서 제공한 2007년 7월 1일부터 2009년 4월 30일까지의 도체등급판 정자료 20,450,773두의 등지방두께, 도체중 및 육질등급 자료를 이용하였으며, 등지방두께와 도체중 에 대한 환경효과를 추정하였고, 성별, 등지방두께별, 도체중별 등급출현율을 추정하였다. 조사된 형 질의 평균치는 도체중과 등지방두께가 각각 85.97±0.002 kg, 20.76±0.001 ㎜로 나타났다. 성의 효과 에서는 거세돼지의 도체중과 등지방두께가 각각 86.25±0.003 kg과 22.55±0.002 ㎜로 나타나, 다른 암 퇘지나 수퇘지에 비하여 유의적으로 높게 나타났고, 도축 년도의 효과에서는 년도가 경과함에 따라 도체중과 등지방두께가 증가하는 경향을 나타내었으며, 도축 계절의 효과에서는 도체중의 경우 겨 울에 87.00± 0.007 kg으로 유의적으로 무겁게 나타났고, 등지방두께의 경우 가을에 19.32±0.004 ㎜로 유의적으로 두껍게 나타났다. 성에 대한 육질등급 출현율의 경우 온도체와 냉도체 모두 거세돼지의 상위등급 출현율이 암퇘지에 비하여 높게 나타났다. 도체중에 대한 육질등급 출현율의 경우, 온도체 와 냉도체 모두 84∼88 kg 그룹에서 상위등급 출현율이 다른 그룹에 비하여 높게 나타났다. 등지방 두께에 대한 육질등급 출현율의 경우 온도체와 냉도체 모두 22∼24 ㎜ 그룹에서 상위등급 출현율이 다른 그룹에 비하여 높게 나타났다.

Salivary lipocalin (SAL1) is a member of the lipocalin protein family that has a property to associate with many lipophilic molecules and was identified as pheromone-binding protein in pigs. Our previous study has shown that SAL1 is expressed in the uterine endometrium in a cell type- and implantation stage-specific manner and secreted into the uterine lumen in pigs. However, function of SAL1 in the uterus during pregnancy in pigs is still not known. To understand physiological function of SAL1 in the uterine endometrium during pregnancy in pigs, it needs to elucidate the ligand(s) for SAL1. Thus, to identify the ligand for SAL1 in the porcine uterus, we collected uterine luminal fluid from pigs on day 12 of pregnancy by flushing with PBS. Proteins from the uterine luminal fluid were separated by ion exchange chromatography and gel filtration. Fractions containing SAL1 protein were pooled and concentrated. Immunoblot analysis confirmed successful purification of SAL1. Then, we extracted lipids from the purified SAL1 protein and analyzed the lipids by liquid chromatography-mass spectrometry, and predicted to be steroid hormones and prostaglandins as SAL1 ligands. Results in this study showed that SAL1 protein in the uterine secretions has a small lipophilic molecule as a natural ligand. Further characterization of ligand extracted from purified SAL1 will be useful for understanding physiological function of SAL1 during pregnancy and its application to increase the pregnancy rate in pigs.