The ovum pick up(OPU) technique can be used to produce embryos after in vitro culture of ovarian oocytes, can be used for early securement for effective herd early proliferation and excellent Hanwoo genetic resources, It is attracting attention as a very important technique for establishing technology. In addition to in vitro culture techniques, the number of oocytes retrieved depends on the environment and timing of the OPU. This study was conducted to compare and examine seasonal effect to the differences in the number of recovered oocytes, recovery rate and embryo development rate using Korean cattle kept in animal genetic resource research center by OPU technique. The grade of COCs was evaluated by microscopic examination. Grade A had 3 or more layers of cumulus cell and compact cytoplasm. Grade B had 1~3 layers of cumulus cell and compact cytoplasm. Grade C had 1 layers cumulus cell and compact cytoplasm. Grade D was denuded oocyte and poor cytoplasm. The recovery rate was 47.8±3.4% in summer (June to August) and 51.6±3.8% in autumn (September to October). The number of oocytes was 5.7±0.6 in summer and 5.2±0.7 in autumn. Oocyte grade A and B was 46.2%±6.3% in summer and 51.1±5.0% in autumn. The cleavage rate was 46.1±7.1% in summer and 65.0±11.3% in autumn. Blastocyst development rate was 19.9±9.4% in summer and 29.0±8.7% in autumn. There was no difference the recovery rate of oocytes and the number of embryos between summer and autumn. Cleavage rate and blastocyst rate of autumn was higher than summer. we will investigate to study the appropriate method for the production of Hanwoo embryos and the systematic comparison.

To test the muscle cell specific gene expression, we examined the ability of human α-skeletal muscle actin (ACTA) promoter or human myoglobin (hMb) promoter to direct the expression of the GFP gene in both muscle and non-muscle cells, respectively. C2C12 cells, a mouse myoblast cell line, provide a powerful model to study skeletal muscle differentiation in vitro. We intended to use this cell line as a model for skeletal muscle-specific gene expression during myogenic differentiation from myoblast to myotubes. We compared marker gene expression profiles of proliferating and differentiated C2C12 cells using RT-PCR and fluorescent microscopy analysis. Also, we found that the expression of PCK1 gene under the control of ACTA promoter was proportionally increased as C2C12 differentiated into myotube form. PCK1 is involved in the regulation of gluconeogenesis. In previous research, transgenic mice with overexpressing PCK1 in skeletal muscle showed a greatly enhanced level of physical activity, which extends well into old age. This is due, in part, to an increased number of mitochondria and a high concentration of triglyceride in their skeletal muscles. These mice also had very little body fat, despite eating 60% more than controls. We also constructed a mesenchymal stem cell line and fetal fibroblast cell line for the experiments aiming to make transgenic animals in which the PCK1 gene is specifically expressed in muscle tissue. Accumulated knowledge of this approach could be applicable to a variety of related biological areas including transgenic animal research, gene function study, anti-aging study, etc. This work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Export Promotion Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (316002-5).

The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each 12.58 ± 8.31 and 13.25 ± 7.86. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen (3.75 ± 1.98 vs. 8.23 ± 6.07, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

Cryopreservation of bovine embryos is used to efficiently implant surrogate mothers. It has been widely accepted that high lipid content in the oocyte interrupts its survival during freeze-thaw cycles. Serum component in the culture medium is thought to increase the embryo`s lipid contents. Conversely, L-carnitine stimulates lipid metabolism by transporting long chain fatty acids into the mitochondria. Objective of this study was to analyze the effect of L-carnitine supplementation in IVM medium and defined IVC medium on the development, lipid contents and the cryosurvival of bovine IVF embryos. 0.0, 1.5, 3.0 and 6.0 mM L-carnitine was supplemented in IVM medium, respectively (IVM-LC 0.0, LC 1.5, LC 3.0 and LC 6.0). Development rate from the 2cell to the morula stages was higher in IVM-LC 3.0 groups than those of IVM-LC 6.0 (p<0.05). But there were no significant differences among the other groups in the blastocyst rates and lipid content results. When 0.0, 1.5, 3.0 and 6.0 mM L-carnitine were supplemented in IVC medium (IVC-LC 0.0, LC 1.5, LC 3.0 and LC 6.0), development competence was not significantly different between those embryos. Lipid contents of embryos treated L-carnitine (IVC-LC 1.5, 3.0 and 6.0) were significantly lower than embryos of non-treated group. L-carnitine was supplemented 0.0, 1.5, 3.0, 6.0 mM during IVM and 3.0 mM during IVC (LC 0.0 - 3.0, LC 1.5 – 3.0, LC 3.0 – 3.0, LC 6.0 – 3.0) and cryosurvival of blastocysts confirmed after freezing-thawing. There were no significant differences on development, but LC 3.0 – 3.0 was significantly lower lipid contents than other groups. And LC 3.0 – 3.0 had better survival rates and hatched rates of blastocysts than LC 0.0 – 0.0. In conclusion, supplementation of L-carnitine in defined IVC medium decreases lipid contents. And L-carnitine supplementation improves cryosurvival and developmental ability of bovine IVF embryos.

The early-onset familial Alzheimer's disease (EOFAD/ FAD), the less common type of Alzheimer's disease (AD) currently affects a vast number of individuals worldwide. This type is being inherited as an autosomal dominant fashion. Missense mutations on Amyloid precursor protein (APP) and Presenilins 1 and 2 (PSEN1 & PSEN2) are known as major genetic factors in FAD. Conversely, missense mutations on microtubule-associated protein tau (MAPT) are also thought to involve. Up to date, several triple-transgenic animal models with muted forms of the human APP, PSENs and MAPT have been reported. Compared to other animals, canines are more emotional and their disease signs can be easily diagnosed. This attempt was to develop a triple transgenic canine model for the AD. We have obtained the coding sequences of APP, PSEN1 and MAPT from Dana-Farber/Harvard Cancer Center DNA resource core at HMS and incorporated several common AD mutations. The transgenic construct is composed of hNSE (ENO2) promoter-driven three AD genes fused together with modified 2A sequences. It was transfected into the canine fetal fibroblasts which were then used to perform somatic cell nuclear transfer (SCNT). The viable transgenic embryos were obtained after in vitro culture and the GFP was detected. In this study, we have successfully produced viable triple transgenic canine cloned embryos using SCNT technique. These transgenic canine embryos will be further developed into canines with FAD. The transgenic canines will be a good candidate in the AD research field.

Epizootic HPAIV, H5N6, and H5N8 infections produced severe loss in poultry and wild birds in the Republic of Korea from 2016 to 2017. But pathological lesions and antigen distribution of the novel HPAIV H5N6 clade 2.3.4.4 in natural cases have been rarely reported. Herein, we describe the pathological lesions and antigen localization in chickens (layer and Korean native), ducks, and Japanese quail naturally infected by HPAIV H5N6. Grossly, severe reddening, swelling, and some necrotic foci, which were similar to septicemia or viremia, were observed in skin and many visceral organs including trachea, lung, liver, spleen, and pancreas. Histopathologically, pulmonary congestion and edema, as well as necrotizing hepatitis, splenitis, pancreatitis, myocarditis, and encephalitis were observed. Immunohistochemically, numerous HPAIV antigens were detected in necrotic parenchymal cells and in blood vessels of the respiratory, lymphoid, digestive, urinary, nervous, and cardiovascular systems. The results indicate that HPAIV H5N6 spread to the entire body via blood and caused severe damage throughout the entire body. The HPAI H5N6 clade 2.3.4.4 virus was isolated from samples of all four cases.

In order to develop the D. adamsi breeding technique that is highly likely to be used as an emotional pet insect, the results of the D. adamsi was collected in the large exhibition Jang Tae-san in Daejeon to the indoor breeding began to spawn after 10 days after the training, the egg laying period was 33.6 days, the average number of spawn per female was 27.7, and the period was 11.8 days. D. adamsi larvae bred with food of oak-boiled molted two times, the duration of 1st larvae development was 11.8 days, 2nd larvae 14.5 days, and the third larvae was 29.4, and the larvae were used to build houses using Masato(soil) and in the pupae, and then in April of the following year. D. adamsi has a characteristic to build a house with fallen leaves and spawn one or two eggs in the fallen leaves, and the results of the spawn mat test using fallen leaves, leaf mold and a sawdust, etc., and the number of spawn was high in the Masato was spread about 3 ~ 5cm, and a fallen leaves on a 10 cm or so, and the growth and survival rate of the larvae were also high. Larvae breeding density was 2 ~ 3 ℓ in the container size to raise the object was normal growth, the higher the density mortality was high and the growth was sluggish. Larvae feeding conditions were normally developed in dry fallen leaves and fermented fallen leaves, compared to the sterile fallen leaves, oak sawdust, sterile fallen leaves and oak sawdust was abnormal, the mortality rate was higher than 50%.

Insect-killing fungi have high potential in pest management. A deeper insight into the fungal genes at the whole genome level is necessary to understand the inter-species or intra-species genetic diversity of fungal genes, and to select excellent isolates. In this work, we conducted a whole genome sequencing of Beauveria bassiana (Bb) JEF-007 and characterized pathogenesis-related features and compared with other isolates including Bb ARSEF2860. A large number of Bb JEF-007 genes showed high identity with Bb ARSEF2860, but some genes showed moderate or low identity. The two Bb isolates showed a significant difference in vegetative growth, antibiotic-susceptibility, and virulence against Tenebrio molitor larvae. When highly identical genes between the two Bb isolates were subjected to real-time PCR, their transcription levels were different, particularly in heat shock protein 30 (hsp30) gene which is related to conidial thermotolerance. In several B. bassiana isolates, chitinases and trypsin-like protease genes involved in pathogenesis were highly conserved, but other genes showed noticeable sequence variation within the same species. Given the transcriptional and genetic diversity in B. bassiana, a selection of virulent isolates with industrial advantages is a pre-requisite, and this genetic approach could support the development of excellent biopesticides with intellectual property protection.

최근 기온이 높아지면서 강원도 대관령 감자 재배지로 날아오는 진딧물 발생이 늘어나고 있다. 그 중 복숭아혹진딧물(Myzus persicae)은 전 세계적으로 분포하고 경제적으로 중요한 해충으로, 감자, 고추 등 노지작물에 흡즙을 통한 바이러스 매개하고 생육저하를 일으키는 피해를 주고 있다. 본 연구는 복숭아혹진딧물의 지리적 차이에 따른 집단의 유전적 관계 규명을 통해 기주식물, 메타집단 등의 군집구조 및 발생패턴을 분석하고, 이러한 요인으로 인한 진딧물 집단의 분산 및 이주 관계를 추정하였다. 최종적으로 개체들 간의 집단유전학적 관계규명을 통해 이들의 확산과 비래 양상을 규명하고자 한다.

Cymbidium flower is mainly grown for exportation to China and Japan, but detection of a few pests including the cotton aphid, Aphis gossypii (Hemiptera: Aphidae), necessitated post-harvest treatment for casual exportation. Thus, we irradiated electron beam to cotton aphids occurring in cymbidium to establish post-harvest method for casual exportation of the flowers. For cymbidium, six categories of product quality were examined after eight different doses were irradiated (100, 200, 300, 400, 500, 600, 800 and 1,000 Gy). One thousand Gy to cymbidium caused an extreme deterioration only in vase life in both colors compared to control (0 Gy). In the case of cotton aphids, adult longevity decreased from 11.23 (100 Gy) to 4.70 (400 Gy) when four different doses were irradiated (100, 200, 300, and 400 Gy), with control being an average of 20.89 days. The numbers of total first instar nymph produced per adult was not significantly differed among five doses (2.21 ~ 2.74 individuals), but was substantially lower compared to control (an average of 51.46; P < 0.0001). Live F1 nymphs did not develope to adults at all five doses, except for a single nymph at 100 Gy, which was dead right after emergence. The results of probit analysis indicated that majority of adults required 3.33 ~ 7.55 days for 90% mortality at 200 Gy and higher, but at 100 Gy it required 41.56 days. Therefore, higher than 100 Gy might be required for complete control of adult cotton aphids and their F1.

This study investigated the fumigant activity of phosphine (PH3) on 2 kinds of mealybug (Pseudococcus longispinus and P. orchidicola (Hemiptera: Pseudococcidae)) adults and nymphs. All of the two mealybugs adults showed higher LCT99 values than nymphs, and P. longispinus showed higher tolerance than P. orchidicola in a 12 L desiccator. The absorption of phosphine on 13 nursery plants showed 12.2~41.5% difference depending on the plant. All of the mealybugs treated with phosphine 2 mg/L in 0.5 m3 fumigation chamber for 4 h showed 100% fumigant activity, except P. longispinus adult (approximately 90% at bottom part). However, when the exposure time was increased to 24 h, all of them showed 100% mortality. In the treatment of 10 m3 container, the 24 h treatment of phosphine showed 100% mortality to P. longispinus and P. orchidicola adults and nymphs. In all the experiments, no phytotoxicity of phosphine observed on 13 plants until 1 month after treatment.

Whitefly (Hemiptera: Aleyrodidae) pests, including the tobacco whitefly, Bemisia tabaci, and the greenhouse whitefly, Trialeurodes vaporariorum, are economically important in agriculture. With the annual growth of the domestic fresh fruit export market, various quarantine treatment methods are being used to export strawberries of better quality. The objective of the present study was to evaluate the effects of gamma rays on the development and reproductive sterility of B. tabaci and T. vaporariorum. In both species, the eggs were completely inhibited from hatching at 50 Gy, and the emergence of 3rd instar nymphs was completely suppressed at 150 Gy. Some adult B. tabaci and T. vaporariorum spawning occurred at 100 Gy and 70 Gy, respectively; however, at these irradiation levels, F1 hatchability was completely inhibited. Dosimetry results showed that the penetrating power of gamma ray in the strawberry-filled box was the lowest at the mid-box position. Therefore, B. tabaci and T. vaporariorum were placed in the middle of the strawberry-filled box and irradiated. A gamma-ray irradiation of 100 Gy suppressed the development and reproduction of eggs and adults in both B. tabaci and T. vaporariorum. However, the development of 3rd instar nymphs was inhibited at 300 Gy for B. tabaci and 200 Gy for T. vaporariorum. Our data suggest that at least 300 Gy should be used for the control of these two types of whitefly for strawberry export.