The present study was undertaken to determine the optimal condition for parthenogenetic activation of rabbit oocytes by electric stimulation in vitro in an attempt to develop nuclear transplantation techniques for cloning mammalian embryos and animals. Freshly ovulated oocytes were collected from superovulated rabbits from 13 to 26 hrs. after hCG injection. The cumulus-free oocytes were activated parthenogetically by repeated stimuli of square direct electric pulses in O.3M mannitol solution. After applying electric stimulations of different voltages, pulse durations and pulse times, all of the oocytes were cultured in TCM-199 with 10% FCS for 96 hours in a 5% incubator, and their developmental potential in vitro was examined. The higher activation rate (68.9%) was achieved at the voltage of 2.0kv/cm, the pulse duration of 60 sec and three pulse times and the activation rate of 100% was achieved at the pulse duration of 100 and 200 sec, the voltage of 1.5kv/cm and three pulse times. Although the higher rates of activation of oocytes were achieved at 100 and 200 sec, none of them developed to blastocyst in vitro. The oocytes collected 18~20 hours post hCG injection showed the highest rate of activation and development to blastocyst in vitro than the oocytes collected 13~15 or 25~26 hours post hCG injection. Therefore, it can be suggested that the application of electric stimulation of 2.0kv/cm, 60 sec and three pulse times to the oocytes collected at 18~20 hours post hCG injection would be more beneficial for the parthenogenetic activation of oocytes in rabbits.

Nuclear transplantation techinque has been found to be the most potential and efficient method for producing a large number of genetically identical animals from a single embryo. The technical development of nuclear transplantation in mammals and its application to the production of cloned animals were reviewed. For the efficient and successful production of cloned embryos by nuclear transplantation, selection and micromanipulation of recipient eggs or embryos as capacious recipient cytoplasm, and benefitial preparation of multiple totipotent embryonic cells as donor nuclei, and also fusion technique are very critical. Recent works approaching to these critical points were introduced and discussed.

Nuclear transplantation technique is known as the most potential and efficient method for producing a large number of genetically identical animals from a single embryo. The technical development of nuclear transplantation in mammals and its application to the production of cloned animals are described. For the efficient and successful production of cloned embryos by nuclear transplantation, the right selection and micromanipulation of recipient eggs or embryos as capacious recipient cytoplasm, the adequate and benefitlal preparation of multiple totipotent embryonic cells as donor nuclei, and also the fusion technique are very critical. Recent studies approaching to these critical points are introduced and discussed. Up to date, the overall efficiency of production of cloned embryos and offspring in livestock is estimated to be low. Further technical development of nuclear transplantation will enable large-scale production of cloned livestock and in near future the commercial cloning of animals will become a reality.

Abnormalities of structure and morphology of chromosomes concentrated with genetic materials, DNA, are directly related to phenotypical performances of animals. So, cytogenetical research in domestic animals is important to prevent congenital deformity and improve genetic performances. Especially utilities of egg transfer technique combined with cytogenetical study can be accelerated by the wide spread of the best genetic sources dependent on the micromanipulation and sexing of eggs.

This paper reviews the most important steps that have generated consistent progress in principles and developmental progress of embryo cryopreservation, and also study on freezing procedure and its application by conventional method and current improved method for freezing procedure and its appilcation of embryo cryopreservation in farm animals. Four were of particular interest: 1.The transport of water across the ccli membrane (zona pellucida) during freezing and thawing accordinglyplays a role in determing whether the celi survives. This movement of water is controlied mainly by extracellular phase changes and by the nature and concentration of any cryoprotective agent present. Therates of cooling, freezing and warming, and the intervals over which they are applied are further decisi've factors in determining whether a cryopreservation procedure allows survival after thawing. 2.The first successful deep freezing experiments with sheep morula and blastocysts during the seventies were based on the early procedures used for mouse embryos.Current research during the eighties is developed with the aim of simplifying and improving current procedures such as one-step dilution and rapid or ultra-rapid cooling by using the model of laboratory animals. 3.The conventional method for the embryo cryopreservation is described. An alternative to this method which may result in high survival and also in reducing of the freezing and thawing time is done by combing a permeable cryoprotectant such as glycerol, DMSO or propanediol and a non-permeable compound such as sucrose, trehalose, raffinose or lactose. 4.Finally a different approach to the preservation of embryos, named vitrification, is introduced. This procedure depends upon the ability of concentrated solutions of cryoprotective agents such as glycerol and propanediol to supercool to very low temperature (-196) during rapid cooling before solidifying without formation of ice. However, more complete data are necessary for successful vitrification of blastocysts.

해양에 대한 인간의 이용과 활동의 증가는 해양오염, 해양생태계 및 서식지 파괴로 이어지고 있다. 이러한 결과, 특히 생애주기가 길고 개체수가 적은 해양동물은 멸종위기에 내몰리고 있는 실정이다. 따라서, 우리나라 정부는 2010년부터 수족관이나 해양박물관과 같이 인 공적인 서식장소 11개를 서식지외 보전기관으로 지정하여 멸종위기에 처한 해양생물들을 보전하기 위해 노력하고 있다. 그러나, 해양생물의 보전 서비스를 제공하는 서식지외 보전기관에 대한 중요성과 그 경제적 가치를 추정한 연구는 아직 이루어지지 않고 있다. 따라서 본 연구의 목적은 우리나라 해양동물의 보호와 증식을 위한 관리방안인 서식지외 보전기관의 운영정책에 대한 일반시민들의 정량적 지지도를 추정하는 것이다. 이를 위해 우리나라 푸른바다거북의 보전을 위해 시식지외 보전기관으로 운영되고 국립해양박물관의 해양동물 보전역할에 대한 경제 적 가치를 추정하였다. 연구의 결과, 서식지외 보전기관과 같은 비시장재인 공공재에 대한 대표적인 가치추정법인 조건부가치추정법을 적용 하여 추정된 서식지외 보전기관(국립해양박물관)의 경제적 가치는 약 418억원에서 최대 약 781억원으로 나타났다. 이러한 본 연구의 결과는 해양동물관리 정책자들에게 우리나라 연안해역에 서식하는 멸종위기에 처한 해양동물의 효율적인 관리방안 수립에 유용한 기초자료로 활용 될 수 있을 것이다.

Genome editing that allows targeted mutagenesis in higher eukaryotic cells and organisms is broadly useful in biology, biotechnology, and medicine. We have developed zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and Cas9 RNA-guided engineered nucleases (RGENs), derived from the type II CRISPR/Cas prokaryotic adaptive immune system, to cleave chromosomal DNA in a targeted manner, producing DNA double-strand breaks in cells, the repair of which via endogenous systems gives rise to targeted genome modifications. The Cas9 protein, when complexed with small guide RNAs (sgRNAs), recognizes and cleaves target DNA sequences complementary to the guide RNAs in vivo, inducing targeted genome modifications at high frequencies in cultured cells and whole organisms. Despite broad interest in RNA-guided genome editing, RGENs are limited by off-target mutations. Here, we show that off-target effects of RGENs can be reduced below the detection limits of deep sequencing by choosing unique target sequences in the genome and modifying both guide RNA and Cas9. Furthermore, we deliver purified recombinant Cas9 protein complexed with sgRNAs (RGEN ribonucleoproteins (RNPs)) to animal embryos and cultured human cells including hard-to-transfect pluripotent stem cells to achieve highly efficient RNA-guided genome editing in cells and whole organisms. RGEN RNPs cleave chromosomal DNA almost immediately after delivery and are degraded rapidly in cells, reducing off-target effects and mosaicism.

Infectious diseases in domestic animals of carcase were increasing every year. According to the monthly report for infectious disease from Ministry for Food, Agriculture, Forestry and Fisheries, 11,954 heads of cattle, 518,178 heads of pig, 232,850,244 heads of bird were reported in 2011. Infectious diseases of carcase almost spread on a national scale. Dead or emergency preventive carcase are completely treated on bury. Since the treatment of carcase on bury are generating soil, underground water and source water pollution, introduction of new preventive system should be considered. In this study, literature survey and case studies to prevent the spreading of virus for infectious disease and second environmental pollution were investigated. The actual experiments using the existing incineration facilities were also performed to ensure the possibilities of safe treatment of carcase. On the other hand, the moving-type incineration are also being developed and its operation manual will be prepared. Among the investigated incinerators, stoker type incinerator was not suitable for the treatment of carcase, however, other incinerators such as fluidized-bed type incinerator, rotary kiln type incinerator ware shown to be suitable. But even for the stoker type incinerator, if the pre-treatment facilities( grinding, crushing) are installed, it will also be a suitable method. The analytical results for air pollutants(including dioxin) emitted from the final exit were all satisfied to the air pollution emission standards.

As increasing incidents of FMD (Foot and Mouth Disease) in recent years, the country has struggled with huge economic losses and environmental problems. Because of relying only on the burial method according to domestic condition, it needs to consider the alternative measure such as the incineration with being no secondary environmental pollution. In addition, such FMD and AI (Avian Influenza) as classifying in the first-class malignant diseases are very important to be ready with rapidly initial response because of the fact being quickly spreaded with high infection speed. Accordingly, a favorable initial response by the introduction of mobile incinerators has been forced to consider. In this study, it analyzed and compared the existing disposal regulations and methods of carcasses to establish the reasonableness about introducing an incineration technology. In addition, domestic and international disposal status was compared as investigating regulations or disposal law, guidelines of livestock in major developed countries. To introduce the mobile incineration facilities in domestic, it is surveyed international examples and related regulations of using and developing mobile incinerators. The results of study could be used as basic information to design and utilize a mobile incineration process for slaughter animals by deceases.