돈분뇨 액비 시용 후 경운처리에 의한 청보리, 호밀, 트리티케일, IRG의 생육과 잡초발생량 차이를 구명하였다. 파종기에 한발이 지속된 포장의 수분함량을 조사한 결과 화학비료 처리구와 무처리구에서는 8.5 ~ 9.1%를 나타냈으나, 액비를 살포한 포장은 20.5 ~ 23.1% 수준이었다. 파종 15일 후에 유묘출현율은 액비 처리구에서는 화학비료 및 무처리구에 비하여 청보리는 2.9배, 호밀과 트리티케일은 각각 2.2배, 2.4배 높게 나타났다. 또한 시험작물 모두 초장이 액비시용구에서 약 2배 이상 증가하였다. 월동 후 최고분얼기에는 모든 작물의 초장이 화학비료 및 무처리구에 비하여 액비처리구가 1.1 ~ 1.3배 길었고 경수도 비슷한 경향을 보였으며, 분얼수는 청보리, 호밀 및 트리티케일이 각각 4.5개, 5개 및 6개/개체로 화학처리구에 비해서 1개체씩의 증가를 보여 한발에 의한 초기 생육차이가 월동 후에도 지속되는 것으로 확인되었다. 한편 잡초발생은 답리작포장이기 때문에 뚝새풀이 주로 발견되었으며, 액비 처리구는 경운으로 m2당 잡초발생수가 5개 수준이었으나, 화학비료 처리구는 무경운으로 145개 발생되어 29배의 잡초발생 차이를 보였다. 월동 후 액비시용하고 경운을 하면 비료효과가 지속되어 화학비료 처리구보다 우수한 생육을 보였으나, 액비시용 후 무경운 처리는 낮은 생육을 보였다. 수확기 생체중 역시 액비처리후 경운처리구가 가장 높았으며, 청보리, 호밀, 트리티케일, IRG 모두 증수되었다. 따라서 한발이 지속될 경우 액비살포 후 경운은 작물의 영양원으로 뿐만 아니라, 수분공급의 효과로 종자의 발아를 촉진시켜 초기생장 및 수량증수에 크게 기여하며 부수적으로 잡초발생도 감소시키는 것으로 판단된다.

The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

새만금 간척지에서 돈분퇴비를 이용하여 수수×수단그라스 교잡종 재배 후 토양변화에 수량성을 평가하기 위한 시험연구 결과는 아래와 같았다. 시험토양은 용적밀도가 높고 공극률이 낮은 간척지토양으로돈분퇴비 시용 후 수수×수단그라스를 재배한 결과 토양경도는 1.2~3 mm까지 증가하였으며 용적밀도는 0.14~0.17Mg·m−3감소하였으나, 이는 토양수분 등 계절적인 요인에 기인한 것으로 생각된다. 토양물리성 중 토양경도와 용적밀도의 관행시비와 돈분퇴비 시용구간의 차이는 없었다. 토양pH, 토양유기물 및 유효인산은 관행시비와 돈분퇴비시용구 처리간 차이가 뚜렷하였으며, 특히 토양유기물과 유효인산은 돈분퇴비의 투입량이 증가할수록 토양 중 함유량의 증가가 뚜렷하였다. 수수×수단그라스 교잡종 건물수량은 가축분퇴비구에서는 질소의 총투입량이 증가할수록 수량이 증가하였다. 새만금 간척지내에서 돈분퇴비를 이용하여 수수×수단그라스 교잡종 재배시 돈분퇴비를 기비로 ha당 17,210 kg이상 시용하면 관행시비와 비슷한 수량이 생산되었다.

The objective of this study was to examine the effect of in vitro culture media on embryonic development of in vitro-matured (IVM) oocytes after parthenogenetic activation (PA) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 22~26 h. IVM oocytes were activated by electric pulses and cultured in porcine zygote medium-3 (PZM-3) and North Carolina State University-23 supplemented with essential and non-essential amino acids (NCSU-23aa). These media were further modified by supplementing 2.77 mM myo-inositol, 0.34 mM trisodium citrate, and -mercaptoethanol (designated as mPZM-3 and mNCSU-23aa, respectively). Culture of PA embryos in mPZM-3 significantly increased development to the blastocyst stage than culture in NCSU-23aa (36.2% vs. 24.8%, p<0.05). Modified PZM-3 showed a significantly higher blastocyst formation than NCSU-23aa in both groups of embryos that were activated at 44 h and 48 h of IVM (51.0% vs. 35.5% and 49.0% vs. 34.2% in oocytes activated at 44 h and 48 h of IVM, respectively). Irrespective of the follicle diameter where oocytes were collected, embryonic development to the blastocyst stage was increased (p<0.05) by the culture in mPZM-3 compared to culture in NCSU-23aa (25.9% vs. 34.2% and 32.9% vs. 44.8% in embryos derived from small and medium size follicles, respectively). Our results demonstrated that culture media had significant effect on preimplantation development PA embryos and that mPZM-3 was superior to mNCSU-23 in supporting development to the blastocyst stage in pigs. This beneficial effect of mPZM-3 on embryonic development was not impaired by other factors such as time of oocyte activation and origin of immature oocytes (small and medium size follicles).

Miniature pig sperm cryopreservation is continually researched in biotechnology for breed conservation and reproduction. It is important to control the temperature at each stage of cryopreservation and cryoprotectant. It is also necessary to find the optimal cryoprotectant concentration and chemical elements of the extender. Recently, many studies have used various cryoprotectant materials, such as dimethyl sulphoxide (DMSO), ethylene glycol (EG), antifreeze protein (AFP), amides, and glycerol. Glycerol is a commonly used cryoprotectant. However, glycerol has critical cytotoxic properties, including osmotic pressure and it can cause irreversible damage to live cells. Therefore, We focused on membrane fluidity modifications can reduce cell damage from freezing and thawing procedures and evaluated on the positive effects of trehalose to the viability, chromatin integrity, and motility of boar sperm. Miniature pig sperm was separated from semen by washing with modified- Modena B (mMB) extender. After centrifugation, the pellet was diluted with the prepared first extender. This experiment was designed to compare the effects that sperm cryopreservation using two different extenders has on sperm chromatin. The control group used the glycerol only and it was compared with the glycerol and glycerol plus trehalose extender. Sperm viability and motility were evaluated using WST1 assays and computer-assisted semen assays (CASA). Chromatin structure was examined using acridine orange staining. For the motility descriptors, trehalose caused a significant (p<0.01) increase in total motility ( in glycerol vs. in glycerol + trehalose) and progressive ( in glycerol vs. in glycerol + trehalose). A significant (p<0.05) increase in VAP ( vs. ), VSL ( vs. ), VCL ( vs. ), STR ( vs. ), and LIN ( vs. ) were also detected, respectively. The sperm DNA fragmentation index was 48.8% to glycerol only and 30.6% to glycerol plus trehalose. Trehalose added group showed higher percentages of sperm motility, stability of chromatin structure than glycerol only. In this study, we suggest that trehalose is effective in reducing freezing damage to miniature pig sperm and can reduce chromatin damage during cryopreservation.

The purpose of this study is to investigate concentration and emission coefficients of 22 odorous compounds, which are regulated by the domestic act, emitted from pig buildings by on-site survey. The odorous compounds which were detected in at least one pig building were ammonia, hydrogen sulfide, methyl mercaptan, dimethyl sulfide, dimethyl disulfide, trimethyl amine, stylene, toluene, xylene and methyl ethyl ketone whereas other 12 odorous compounds were not detected in pig buildings. In general, indoor concentrations of odorous compounds in pig buildings were higher in scraper type than slurry type based on pig manure collection system and higher in enclosed type than winch-curtain type based on ventilation mode, respectively. In monthly distribution of odorous compounds, their concentrations in September and October when ventilation rate in pig building decreased relatively were generally higher than those in July and August when ventilation rate in pig building is relatively high. On the contrary, the emission coefficients of odorous compounds in pig building were generally higher in July and August than September and October. The levels of emission coefficients of odorous compounds obtained from this study were similar or slightly higher compared to those reported previously from foreign countries.

Hyperacture rejection (HAR) of pig organs, upon xenotransplantation into primates, could partly be overcome by knocking out the alpha-Gal gene. However, xenotransplanted organs may still undergo immunological acture rejection (AR) or acute vascular rejection (AVR). Among several genes involved in AR and AVR, the hCD47 evades the monocyte/ macrophage mediated phagocytosis by identifying the self/non-self signal (CD47-SIRPa) whereas hTFPI participates in the regulation of coagulation pathway by acting upstream of the thrombin. In this study, we investigated hCD47 and hTFPI as two possible candidates for avoiding AR and AVR, respectively upon pig-to-human xenotransplantation. A co-expression vector for hCD47 and hTFPI was constructed using 2A peptides system (F2A) and transfected into the porcine kidney cell line (PK-15). The transfected cells stably expressed both hCD47 and hTFPI mRNA and proteins. Co-culture of non-transfected, hCD47-transfected, hTFPI-transfected or hCD47+hTPFI-transfected PK15 cells with natural killer (NK) cells, monocytes and macrophages confirmed the cytotoxic effect of hCD47 and revealed a synergistic effect of hCD47 and hTFPI co-transfection. There was an attractive survivability of 25～30% on each type of innate immune cell, NK cell and macrophage. These results suggest that transgenic pigs, genetically modified for hCD47 and hTFPI may be useful for overcoming xenograft rejection. Furthermore, cotransfection with hTFPI may enhance the cytotoxic effect of hCD47, possibly by assisting the hCD47-SIRPa binding by an unknown mechanism.