True hermaphrodites are animals of equivocal sex in which both male and female gonads develop simultaneously. The frequency of true hermaphroditism is higher in pigs than in other domestic animals. Two Korean pigs were diagnosed with true hermaphroditism showing ovotestes, epididymes, penes, and uteri. Histomorphologically, the testicular tissues consisted of Sertoli cells that were devoid of spermatogenic germ cells and showed proliferation of interstitial cells. However, the uteri were of normal architecture and had well-developed uterine endometrial glands. The samples were 38, XX female karyotype without the sex-determining region Y (SRY) gene. The findings of this study could contribute to the understanding of true hermaphroditism in the Korean pig industry. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The aim of this study was to examine the effect of acteoside (the cyclin-dependent kinase inhibitor) on the SCNT efficiency with adult fibroblasts in dog. Canine adult fibroblasts were obtained from muscle and cell cycle of fibroblasts was synchronized by culturing to confluency, serum starvation and treating with 30 μM acteoside for 48 h. Cell cycle stages, cell cytotoxicity (apoptosis) and, prduction of reactive oxygen species (ROS) were analyzed using flow cytometry. The canine cells, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were injected into enucleated in vivo matured oocytes, the couplets were electrical fused and activated by calcium ionomycin. SCNT embryos using acteoside-treated fibroblasts were surgically transferred into oviducts of estrus cycle synchronized recipient dogs. In cell cycle synchronization (G0/G1), there was no significant difference between serum starvations (83.9%) and acteoside treated groups (81.3%) that were higher than confluent group (78.5%). In production of apoptosis, confluent and acteoside treated groups (4.3 and 4.5%, respectively) were generated less than serum starvation group (21.8%). In case of ROS, serum starvation group was induced a significantly higher than other groups. After synchronization of the donor cell cycle, either confluent or acteoside treated, cells were placed with enucleated in vivo-matured dog oocytes, fused by electric stimulation, activated, and transferred into naturally estrus-synchronized surrogates. Fusion and cleavage rate of acteoside treated group were 64.1 and 41.5%, which were higher than those of confluent group (53.9 and 20.6%, respectively). The reconstructed embryo development rates to 4-cell and 8-cell in acteoside treated group were 29.5 and 14.8%, respectively, while confluent group showed 11.1 and 3.2%, respectively. Total 54 SCNT embryos using acteoside-treated fibroblasts were transferred into oviducts of 2 recipient dogs and one recipient finally delivered one puppy, whereas din`t detected pregnancy on transfer of cloned embryos reconstructed with confluent cells in 6 surrogate dogs. In conclusion, the results of the current study demonstrated that canine fibroblasts could be successfully arrested at the G0/G1 stage with reduced the formation of ROS and apoptosis after acteoside treatment. This results may contribute to improve the effi-ciency of canine SCNT. * This research was supported by iPET (Grants 110056-3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Acteoside acts as an anti-oxidative activity and anti-apoptosis in the cells. But, it has been not studied on maturation and development of porcine oocytes. The aims of the present study were to examine the effects of acteoside on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control), 10, 30 and 50 μM. The oocytes maturation rates of groups supplemented with acteoside were no significantly different (81.13, 85.96, 82.95 and 83.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (44.83 vs. 27.75%). And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. In the results. during IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with control oocytes. And reverse transcription polymerase chain reaction (RTPCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes (Mcl-1, Bcl-2 and Bcl-xL), whereas reduced the expression of pro-apoptotic genes (Bax and Bak). In conclusion, based on the results, the effect of acteoside on IVM was not attractive. However, in acteoside treated group, cytoplasmic maturation seemed to be improved with morphologically uniform distribution of cytoplasmic organelles. Furthermore, embryonic development in acteoside treated group was significantly highly increased than that of non-treated group. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes, providing a improved method for porcine oocytes in vitro. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The objective of the current study was to describe in vitro embryo production in Hanwoo, analyzing oocytes yield and embryo production. The effects of oocytes production and the number of OPU procedures per animal on embryo production were also evaluated. OPU was done every 3～4 days during experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, we compared the recovery rate of oocytes based on OPU session (Experiment 1). The average of collected oocytes was calculated from every 10 session. The average number of total oocytes recovered per animalonsessionwas 5.16 (mean). Second, we compared the recovery rate base on collection period of OPU (Experiment 2). The following results show the difference of the number of recovered oocytes in every month during the procedure between the months of session. Every animal shows the constant number of recovered oocytes for the first 5 months. However, the recovery rate of oocytes was decreased from month 6 to 8. Third, we compared the developmental rate to blastocyst in two groups (Experiment 3). Oocytes by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24～48 h after in vitro fertilization (IVF) was 75.8% and blastocyst development rate was 18.8% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (61.1%), blastocyst development rate was higher compared with needle puncture group (28.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio- technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being.

The objective of this study was to evaluate in vitro production of bovine embryos in Hanwoo. Oocytes were collected by ovum pick up (OPU) from ovaries of genetically high-value Hanwoo or by needle puncture from ovaries of slaughtered cattle. OPU was done every 3 4 days duing experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, We compared the in vitro maturation rate in two groups (Experiment 1). 545 oocytes were recoverd from 4 females by 32 trials of OPU and then 433 oocytes were shown MⅡ stage after in vitro maturation (79.4%). In case of needle puncture group, 1905 oocytes were collected and then 1420 oocytes were matured to MⅡ stage during in vitro culture(74.5%). Second, we compared the developmental rate to blastocyst in two groups (Experiment 2). 1420 oocyte by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24 48 h after in vitro fertilization (IVF) was 88.6% and blastocyst development rate was 20.5% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (84.8%), blastocyst development rate was higher compared with needle puncture group (26.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio-technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being. This research was suppoted by Imsil-gun agricultural technology service center.

Historically the rise of contagious diseases and epidemics has been mainly caused by lack of food safety knowledge and negligence in personal hygiene. Food poisoning is an example that have been caused by lack of proper knowledge in food safety and personal hygiene. In order to prove this hypothesis, our team distributed a total of 225 survey sheets to middle school students. Survey questions mainly focused on the students' food safety knowledge, personal hygiene and regular dietary habits. We used 12 items to determine the stages of change in teenagers and divided into 3 stages of change: precontemplation stage (poor problem recognition), contemplation and preparation stage (some problem recognition), and action stage. Out of the 225 students surveyed, 105(47.1%) were male and 118 (52.9%) were female; 44.0% were in 7th grade, 33.0% in 8th grade, and 24.0% in 9th grade. In stages of change model, 3.8% were classified in pre-contemplation stage, 31.1% in preparation stage, and 65.1% in action stage. The awareness of the significance of food safety has a strong positive correlation with the stages of change food safety behavior (P<0.001), which suggesting our questionnaires regarding stages of change and food safety behavior were quite credential. Stages of change were also correlated with the consumption of unsanitary foods (P<0.01) and with the frequent hand washing (P<0.05). The stages of change showed a positive correlation with the food safety knowledge level (P<0.05). The four questions (out of 10) with the least percentage of correct answers were questions about the meaning of Norovirus (23.6%), the meaning of expiration date (43.6%), the meaning of food poisoning (36.4%), and proper food-handling (36.0%).

The leaf beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation of the species we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) collected from seven Korean localities. A total of 18 haplotypes (BARCA01 ~ BARCA18), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 17 sequence types (ITS2CA01 ~ ITS2CA17), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in mitochondrial gene sequence. Phylogenetically, the mitochondrial DNA has shown several haplotypes formed independent groups with substantially high node support (≥ 90%), whereas no such grouping was evidenced for ITS2, indicating different behaviors of the two molecules. Such difference may reflect a diverse dynamics of the species such as biogeographic history, mating behaviors, and also possibly different mode of inheritance of the two molecules, but requires further scrutinized examination of the dataset. In terms of population genetic perspective, overall no population subdivision was detected from both molecules, except for locality 7 (Eocheong islet) from mitochondrial DNA. As more scrutinized analysis is performed, further fruitful inference on the geographic contour of the species might be available.

The silkworm (Bombyx mori), as an industrial insect, possesses a high economic value. Casual discrimination and accumulated genetic information of silkworm varieties are essential ground for the practical utilization and long-term conservation. In this study, nine available microsatellite loci were successfully genotyped from ~50 silkworm strains preserved in Korea. According to genotyping analysis, we obtained 3 ~ 16 alleles per locus, with an average of 7.4, the observed heterozygosity ranging from 0.04 to 0.98, and the polymorphic information content (PIC) ranging from 0.06 to 0.88, revealing that some loci are highly variable. Among 54 strains 13 strains were casually identified by the presence of 17 strain-specific apomorphic alleles. Furthermore, 30 among remaining strains contained strain-specific allele combinations that are also apomorphic to each strain, allowing us to discriminate each of these from other strains by genotyping of multiple loci. These results collectively suggest that the silkworm microsatellite DNA is actually and potentially important molecular marker for the discrimination of the silkworm strains that are preserved as hundreds in Korea, as more loci are genotyped.

The 15,338-bp long complete mitochondrial genome (mitogenome) of the Japanese oak silkmoth, Antheraeayamamai (Lepidoptera: Saturniidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, but differs from the most common type, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. No typical start codon of the A. yamamai COI gene is available. Instead, a tetranucleotide, TTAG, which is found at the beginning context of all sequenced lepidopteran insects was tentatively designated as the start codon for A. yamamai COI gene. Three of the 13 protein-coding genes (PCGs) harbor the incomplete termination codon, T or TA. All tRNAs formed stable stem-and-loop structures, with the exception of tRNASer(AGN), the DHU arm of which formed a simple loop as has been observed in many other metazoan mt tRNASer(AGN). The 334-bp long A+T-rich region is noteworthy in that it harbors tRNA-likestructures, as has also been seen in the A+T-rich regions of other insect mitogenomes. Phylogenetic analyses of the available species of Bombycoidea, Pyraloidea, and Tortricidea bolstered the current morphology-based hypothesis that Bombycoidea and Pyraloidea are monophyletic (Obtectomera). As has been previously suggested, Bombycidae (Bombyxmori and B.mandarina) and Saturniidae (A.yamamai and Caligula boisduvalii) formed a reciprocal monophyletic group.

We determined the complete mitochondrial genome (mitogenome) of the Japanese Oak Silkmoth, Antheraea yamamai (Lepidoptera: Saturniidae) from two overlapping fragments and subsequent shotgun sequencing. The 15,601-bp long A. yamamai mitogenome contains gene arrangement and content identical to the most common arrangement found in lepidopteran insects. Most individual A. yamamai mitochondrial (mt) genes were well within the range found in the respective genes of other insects, except for small ribosomal RNA (1,037 bp). The 336-bp A+T-rich region is relatively smaller than that of other lepidopteran insects. The region is interesting in that it contains tRNA-like structures as found in the A+T-rich regions of other insect mitogenomes. The start codon of A. yamamai COI gene is unusual in that no typical one (ATN) is available. Three of the 13 protein-coding genes have incomplete termination codon T or TA. All tRNA formed stable stem-and-loop structure, except for tRNASer(AGN), the DHU arm of which formed a simple loop as seen in many other metazoan mt tRNASer(AGN).

Bumblebees are important pollinators in greenhouse and have colonized all parts of the World. In Korea, the value of bumblebees is increasing as pollinator. However, the more recent use of reared colonies may ultimately allow pathogens to spread to peripheral areas for bumblebees. Generally, bumblebees are hosts to a large number of parasites which are mites, flies, protozoa, fungi, virus and bacteria. For most of these, very little is known about their effects against host insect, epidemiology or evolutionary ecology. So, we report pathogenic bacteria isolated from Bombus terrestris and B. ignitus at first time in Korea. Bacillus fusiformis and Klebsiella oxytoca are isolated from B. terrestris and confirmed with 16S rRNA gene nucleotide comparison in NCBI genebank. Pathogenicity of B. fusiformis was 35~40% against B. terrestris. Pantoea dispersa and K. oxytoca are isolated from B. ignitus and confirmed with 16S rRNA gene nucleotide comparison in NCBI genebank. Pathogenicity of these species were 35~40% against B. ignitus. These pathogenicity are considered as low-level.

지진하중에 대한 구조물의 동적 거동과 내진성능을 평가하기 위하여 유사동적실험기법이 흔히 사용되고 있으나, 실험장비의 제약과 구조물의 규모 등으로 대부분 축소모형실험에 의존하고 있다. 그러나 일반적인 상사법칙은 탄성범위에서 유도된 것으로 지진거동과 같은 비탄성 거동을 예측하는 경우에는 한계가 있기 때문에 축소모형의 실험결과를 원형 구조물에 직접 적용하는 것은 많은 주의가 필요하다. 본 연구에서는 원형구조물과 축소모형에 대한 철골모형을 실험 대상으로 하여 실제 축소모형 실험결과로부터 원형구조물의 거동을 예측하는 경우의 문제점을 확인하고, 그 해결방법을 모색하고자 한다. 실제로 축소모형실험에서는 원형구조물의 경계조건을 정확히 재현하기 어려우며, 축소모형의 제작과정과 실험과정에서의 모든 오차가 강성의 변화로 반영되어 나타난다. 따라서 본 연구에서는 기하학적 상사율과 축소모형의 변화된 강성비를 함께 고려한 수정된 상사법칙을 제안하였으며, 수정된 상사법칙이 적용된 축소모형에 대하여 유사동적실험을 수행하여 지진응답결과를 원형구조물의 결과와 비교하였다. 축소모형에 대한 유사동적실험결과로부터 제안된 상사법칙을 적용함으로써 원형구조물의 지진응답 재현이 효과적임을 입증하였다.

글루코오즈(GOD) 옥시다제가 고정화된 PVA/키토산 플렌드막과 다공성 폴리아미드 복합막을 통해 인슐린의 투과거동을 살펴보았다. GOD가 고정화된 막을 통한 투과계수는 10-6~10-7cm3/cm2sec이었다. 복합막의 클루코오즈 농도에 대한 변화는 낮은 글루코오즈 농도에서 높았는데 이는 막으로부터 산소의 고갈 때문이었다. PVA/키토산 및 다공성 폴리아미드막을 통한 인슐린의 투과는 글루코오즈 농도에 따라 500mg%까지 점차 증가하였다.

본 연구는 완주에서 재배된 세계 주요 11개 적포도 품종으로 제조된 적포도주의 향기 성분을 headspace-solid phase microextraction 분석법으로 확인하였다. 향기성분은 총 75종이 확인되었다. 아로마화합물은 그들의 OAV 값에 의해 5 그룹으로 나뉘었다. 알콜, 알데하이드, 에스테르, C6 화합물이 11개 적포도주의 주요한 향기성분이었다. Isoamy alcohol 알콜과 phenylethyl 알콜은 11개 포도주에서 공통적으로 꽃향기, 달콤한 향을 나타내는데 중요한 물질이었다. Octanoic acid, ethyl ester, hexanoic acid ethylester은 모든 레드와인에서 과실향과 꽃향, 달콤한 향을 내는 중요한 성분이었다. 1-Hexanol은 모든 포도주에서 분석되었으나 풀향을 나타내는 향으로 나타났다. Chanceller, Malbec, marchel, Nsrsha, Pinot Meunier, Sangiovetto 포도주의 주요 향기성분은 과실 향인 것으로 나타났으며 Cabernet Franc, Cabernet Sauvignon, Sauvignon Vert 포도주의 주요 향기성분은 풀향인 것으로 조사되었다. 또한, MBA와 Narsha 포도주의 경우 꽃향이 주요 향인 것으로 조사되었다. 본 연구 결과를 바탕으로 적포도주용 품종을 육성할 때 선발기준으로 향기성분 분석을 활용할 수 있을 것으로 판단되었다.

Dendropanax morbifera Leveille (Araliaceae) is an endemic species growing in the south-western part of South Korea that has been used in folk medicine and health functional food. In this study, we investigated an extract of quercetin in Jeju D. morbifera by varying different parts (fruit, sprouts, leaves, sprigs, and branches), harvest times, and extraction solvents. In addition, we aimed to establish a simple and reliable HPLC/UV analytical method to determination of quercetin for the quality control and base line data of the Jeju D. morbifera extract as a health functional food ingredient. The analytical specificity was determined with retention time and photo diode array (PDA) spectrum by analyzing quercetin using HPLC and comparing the results to those of extracts. This analytical method for quercetin was validated for its limit of detection (LOD), limit of quantitation (LOQ), precision, and accuracy. A high linearity in the standard calibration curve was obtained, with a coefficient of determination (R2) of 0.9996. Also, the LOD and LOQ values were found to be 0.28 μg/mL and 0.85 μg/mL, respectively, and the recoveries of quantified compounds ranged from 97.91% to 104.10%. Furthermore, the relative standard deviation (RSD) values of data from the intra- and inter-day precision analyses were less than 1.36% and 3.65%, respectively. As a result, the highest quercetin content among the extracts of Jeju D. morbifera leaves was found to be 20.14 mg/g, which was extracted at harvest in May (cultivation period 10 years) with 60% EtOH. All in all, we believe that the results obtained would be helpful in the development of nutraceutics and natural medicines and for the quality control of D. morbifera.

The study compared the juice and wine odorants of the Cheongsoo grape cultivar with those of Chardonnay and Reisling wines. The volatile compounds of the three grape varieties were analyzed using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The most common volatile compounds in the juices from the three cultivars were terpenes, C13-norisoprenoids, etones, alcohols, and aldehydes. Terpenes were established as the most abundant group of volatiles in Cheongsoo grape juice, where as aldehydes predominated in Chardonnay and Riesling juices. Forty-two volatile compounds (acids, alcohols, esters, and others) were detected in the three white wines. The concentration of esters was about four times higher in Cheongsoo wine than in Chardonnay and Riesling wines. Five esters found in the Cheongsoo wine, namely, isoamyl acetate, ethyl butanoate, ethyl hexanoate, ethyl octanoate, and ethyl decanoate, exhibited high odor activity values (OAV) of >1. Furthermore, only Cheongsoo wine had a high OAV for isoamyl acetate odorant, which is associated with banana and sweet aromas. Therefore, the abundant and varied esters are believed to be key volatile fruity/sweet odorants in Cheongsoo wine.

사과는 에틸렌에 의해 호흡량이 일시적으로 상승하는 호흡급등형 과실이다. 에틸렌 발생은 세포벽분해효소 활성화와 세포벽 연화를 진행시켜 사과의 상품성과 저장성을 떨어뜨리는 원인이 된다. 사과의 에틸렌 생합성 과정에는 Md-ACS1 및 Md-ACO1 유전자가 연관되어 있으며, 두 유전자는 과실의 에틸렌 발생량과 경도에 영향을 미치는 것으로 알려져 있다. 본 연구는 사과 국내육성 28품종의 Md-ACS1 및 Md-ACO1 대립유전자형을 분석하고, ‘Fuji(FJ)', ‘RubyS (RS)', ‘Hongro(HR)', ‘Arisoo(AS)', ‘Summer King(SK)', ‘Greenball(GB)', ‘Golden Supreme(GS)'을 대상으로 수확 후 25일까지의 에틸렌 발생량 및 상온저장(20℃) 20일 동안의 경도 연화율을 조사하였다. 그 결과, 낮은 에틸렌 발생량과 관련 된 대립유전자(favorable alleles0(FA)) ACS1-2, ACO1-1이 많을수록 에틸렌 발생량과 경도 연화율이 낮은 경향을 보였다. GS은 ACS1-1/1, ACO1-1/2(FA 1)으로 모든 품종 중 가장 높은 에틸렌 발생 수치와 경도 연화율를 보였다. SK와 GB은 ACS1-1/2, ACO1-1/2(FA 2)으로 ACS1-2/2, ACO1-1/2(FA 3)인 HR와 AS 보다는 높고 GS 보다 낮은 에틸렌 발생량과 경도 연화율을 보였다. ACS1-2/2, ACO1-1/1(FA 4)인 FJ는 RS를 제외한 모든 품종 중에 에틸렌 발생량과 경도 연화율이 가장 낮았다. 본 실험의 결과 Md-ACS1 및 Md-ACO1 대립유전자형과 사과의 에틸렌 발생량 및 경도 연화율은 상관성이 있는 것으로 확인되었다. 따라서, Md-ACS1 및 Md-ACO1 분자표지를 사과 국내육성 품종의 저장성 예측과 육종 효율 향상을 위한 Markerassisted selection에 활용할 수 있을 것으로 생각 된다.

Background : For the GAP standard cultivation, weed occurrence pattern was investigated by planting density and mulching method of Bupleurum falcatum L. Methods and Results : Vinyl and rice straw were used for mulching. The planting density were 10, 20, and 30 cm, and the spacing between plants were 5 ㎝. The amount of weed emergence were examined twice at the end of July and at the end of September. The degree of importance was based on the Braun-Branquet’s dominance value distinction criterion. As a result, the kind of total weeds emerged during the cultivation of Bupleurum falcatum L. were identified as 20 species in 10 families. Amount of weed’s occurrence were the highest in rice straw covering after 10 × 5 ㎝ sowing, and it were lowest in vinyl mulching after 30 × 5 ㎝ sowing. The frequency of weed emergence was highest with 10 species of 8 families in rice straw mulching treatment. On the other hand, the treatment with the least emergence frequency of weeds was 30 × 5 ㎝ sowing after vinyl mulching, and weeds of 7 species of 6 families appeared. In the second survey in September, weeds of 15 species in 9 families were identified in the test field. The amount of weed’s occurrence were the highest in rice straw mulching after 10 × 5 ㎝ sowing, and it were the least in the case of vinyl mulching after 30 × 5 ㎝ sowing. The frequency of emergence of weeds was highest in rice straw mulching after 20 × 5 ㎝ sowing, and the kind of weeds were 11 species of 8 families. The treatment of the lowest emergence frequency of weeds was 30 × 5 ㎝ sowing after vinyl mulching. Three kinds of weeds occurred. Conclusion : From the above results, it is expected that we can suppress the weed occurrence at the planting density of 30 × 5 ㎝ after vinyl mulching at the sowing of Bupleurum falcatum L.