본 연구는 효율적인 RS 조제기술을 규명할 목적으로 실시하였다. 연구는 RS의 저장부위에 따른 발효품질 및 사료성분의 변화에 대하여 검토하였는데, 곤포한 1개월 후에 3개의 RS(bale No. 1, bale No. II 및 bale No. III로부터 부위별 (상층부, 중앙부 및 하층부)로 채취하였다. 건물 함량은 상층부에서 bale No. I, bale No. II 및 bale No. III이 각각 64.5, 87.6 및 로 bale No. II가

본 연구는 도축돈의 난소로부터 난자를 채취하여 체외배양시킨 후 세포 안정제와 원심분리 그리고 OPS를 이용한 유리화동결 하였다. 동결 융해한 수정란을 경산돈에 외과적 또는 비외과적으로 이식하여 자돈을 생산하는 것을 목적으로 수행하였다. 도축돈 난소로부터 채란되어진 돼지 미성숙난은 Funahashi 등(1994) 방법에 따라 체외 성숙-수정-배양하였다. 체외배양액은 glucose-free NCSU 23을 이용하였으며, 5일째에 10% Fatal bovin

This study evaluated the efficiency and compared with different materials of loading vessels for vitrification-plastic/glass, copper grid and nylon. The loading method, vitrification, cryop-reservation and warming method of the oocytes were examined. The loading samples prepared in manual or company-made and sterilized, loaded the COCs selected on each samples and cultured for maturation during 40 hours, and then exposed sequentially to ethylene glycol solution. Thawing method was reversely treated and exposed for warmed oocytes. After oocytes were thawed, fertilized and cultured in vitro for 3-4 hours, rates of development and morphological appearance were examined. The results were as summarized: ㆍOPS from company-made or hand-made of the hematocrit micropipettes, NLS from fishing line and EMG from company-made for EM were used for loading oocytes, respectively. ㆍThe efficiency of freezing method and loading convenience were orderly higher in OPS, NLS and EMG. The optimal capacity per vessel was orderly lowered in NLS, EMG and OPS, respectively. ㆍAfter oocytes were warmed, the recovery rate, morphology and rate of development were orderly higher in OPS, NLS and EMG, respectively. ㆍIn conclusion, OPS has the advantages of achieving a little more survival and preserving results than other two loading methods.

Human embryonic stem (hES) cell lines have been derived from human blastocysts and are expected to have far-reaching applications in regenerative medicine. The objective of this study is to improve freezing method with less cryo-injuries and best survival rates in hES cells by comparing various vitrification conditions. For the vitrifications, ES cells are exposed to the 4 different cryoprotectants, ethylene glycol (EG), 1,2-propanediol (PROH), EG with dime-thylsulfoxide (DMSO) and EG with PROH. We compared to types of vehicles, such as open pulled straw (OPS) or electron microscopic cooper grids (EM grids). Thawed hES cells were dipped into sequentially holding media with 0.2 M sucrose for 1 min, 0.1 M sucrose for 5 min and holding media for 5 min twice and plated onto a fresh feeder layer. Survival rates of vitrified hES cells were assessed by counting of undifferentiated colonies. It shows high survival rates of hES cells frozen with EG and DMSO (60.8％), or EG and PROH(65.8％) on EM grids better than those of OPS, compared to those frozen with EG alone (2.4％) or PROH alone (0％) alone. The hES cells vitrified with EM grid showed relatively constant colony forming efficiency and survival rates, compared to those of unverified hES cells. The vitrified hES cells retained the normal morphology, alkaline phosphates activity, and the expression of SSEA-3 and 4. Through RT-PCR analysis showed Oct-4 gene expression was down-regulated and embryonic germ layer markers were up-regulated in the vitrified hES cells during spontaneous differentiation. These results show that vitrification method by using EM grid supplemented with EG and PROH in hES cells may be most efficient at present to minimize cyto-toxicity and cellular damage derived by ice crystal formation and furthermore may be employed for clinical application.

The advantages of the OPS techniques(Vajta G et al, Mol Reprod Dev 51: 53-58,1998) give 1) high survival rates of various types of eggs, 2) quick and simple process, 3) inexpensive equipment and reduced chilling injury. The efficiency of IVM/IVF technique in the porcine species is relatively lower than that obtained in other species such as ruminants. Two experiments were designed to investigate the effects of in-vitro fertilization of porcine oocytes matures using different OPS protocol for chilling and warming of vitrification. Porcine oocytes from ovaries collected at abattoir were matured for 44 hours in TCM199 Earle's salt supplemental with pyruvate, pff, L-cysteine, hormones and gentamycin. Oocytes were denuded and fertilized with frozen boar semen by common method. Porcine embryos produced routinely by in-vitro culture system of NCSU23 medium. The vitrification and the warming were conducted by OPS method with the glass micropipette instead of straw vessels and modified the protocol of G.Vajta(1999). In Exp 1, Chilling/Warming:Holding Medium(HM)+EG+DMSO/HM +sucrose Medium(SM) at 39 warm stage. In Exp 2, : PBS+CS+EG+Ficoll+ Trehalose/PBS+Trehalose at 25 stage. Filling, freezing, packing, thawing out and further culturing were performed to follow the basic protocol of G Vajta. During IVM-lVC and post-warming, fertilization parameter and developmental potential were compared to and statistically analysed. It was not significantly different from Exp 1 and Exp 2 but 25 of stage was slightly higher on the morula/blastocyst forming rate and better atmosphere for worker than that at 39 stage.

본 실험에서는 볏짚에 사과박의 첨가비율을 달리하여 조제한 사일리지(볏짚:사과박 = 100:0, 80:20, 60:40, 40:60) 혹은 옥수수 사일리지에 건초 및 시판사료의 혼합 비율을 각각 40:40:20로 조제하여 한국 재래산양에게 급여하였고, 이들에 의한 사료섭취량, 소화율 및 질소 축적율 등을 조사하여 농산 가공 부산물의 사료화 가능성을 제시하고자 실시하였으며 그 결과를 요약하면 다음과 같다. 시험사료의 화학적 조성분은 조단백질 함량이 6.3∼

To develop an effective vitrification method, we examined the use of a conventional straw as vessel fur vitrification of mouse oocytes, and to compare the post-thaw survival and chromosome configuration of these oocytes with those vitrified in grids. Intact cumulus-enclosed oocytes were vitrified with DPBS with 5.5 M ethylene glycol and 1.0 M sucrose, and loaded into straws and onto eletron microscopic copper grid fur storing in liquid nitrogen. Intact vitrified and thawed oocytes were karyotying for chromosome. The rates of post-thawed survival were 88.5% in vitrified oocytes with straws, and 83% in vitrified ooctyes with grids. Vitrified and thawed oocytes with straws and grids were increased chromosomal abnormality (31.4% and 30.9%) compared with fresh oocytes (17.8%). The conventional straws can be used as vessel for vitrification to prevent of inflection in liquid nitrogen.

본 연구는 OPS 방법에 의한 돼지미성숙 및 성숙난자의 동결-융해 후 난자의 생존성에 있어서 난구세포의 영향을 검토하였다. 그 결과 미성숙 난자의 동결-융해 후의 성숙율은 난구세포의 부착 (25%) 및 제거 (15%)시 유의적인 차이는 인정되지 않았지만 control group (62%)에 비해서는 유의적으로 낮게 나타났다(P<0.05). 미성숙난자의 동결-융해 후 체외성숙시킨 난자의 체외수정시 난구세포 제거시 (19%), 부착된 (9%) 난자에 비해

This experiment was carried out to determine the effect of storing method and wrapping frequency on the quality of round baled rice straw silage at experimental field of Grassland and Forage Crops Division, National Livestock Research Institute, RDA, Suwo

The quality of the rice straw silage added with apple pomace was investigated in this study and the amount of apple pomace added in different treatments were 0, 20, 40 and 60%, respectively. Crude protein contents (6.4-7.5%) of rice straw silage added wit

본 시험은 원형베일러를 이용한 생볏짚 사일리지의 조제 가능성을 구명하고 적정 첨가제를 선발하기 위하여 1997년 10월부터 1998년 6월까지 축산기술연구소 초지사료과에서 실시하였다. 첨가제는 생볏짚을 원형베일로 만들기전 무처리, 개미산, 당밀, 당밀+요소 및 젖산균제 등 5처리를 두고 난괴법 3반복으로 수행하였으며 사일리지 품질과 가축 섭취량 등을 조사하였다. 조단백질 함량은 당밀+요소 처리구에서 가장 높았으며 소화율은 당밀 및 젖산균 처리구에서 증가하였다. 건물률에 있어서는 개미산 처리구가 가장 높았고 대조구에서 가장 낮게 나타났으며 첨가제 처리간에는 차이가 없었다. 그러나 산도(pH)는 당밀 및 젖산균제 처리구에서 대조구보다 낮게 나타났다. 유기산 함량은 전체적으로 2%이하로 낮게 나타났으며, 젖산 함량은 당밀 및 젖산균제 처리구에서 유의적으로 증가하였고(P<0.05) 낙산 함량은 감소하였다. 암모니아태 질소의 함량은 당밀+요소 처리구에서 유의적으로 높았으며 당밀 및 젖산균제 처리구에서 10% 이하로 낮게 나타났다. 7일간의 가축 급여시험에서는 젖산균제를 처리한 생볏짚 원형베일 사일리지에서 두당 섭취량이 무처리 원형베일 사일리지와 건조볏짚에 비해 높게 나타났다. 생볏짚 원형베일 사일리지 생산비는 수거면적이 넓어짐에 따라 생산비가 크게 낮아졌다. 이상의 결과를 종합하여 볼 때 생볏짚을 원형베일 사일리지로 조제시 당밀 또는 젖산균제를 첨가할 때 가장 우수한 사일리지를 조제할 수 있었고 젖산균제 처리시 가축 섭취량도 증가되는 것으로 나타났다.