We sequenced 17,329 bp of mitochondrial genome (mitogenome) of the black dwarf honey bee, Apis andreniformis (Hymenoptera: Apidae), that lacks ~200 bp of the A+T-rich region for the completion of the genomic sequence. The gene arrangement of A. andreniformis mitogenome is identical to that of A. cerana. However, the genome contains 5 additional tRNALeu(CUN) located 4 copies between tRNAMet and tRNAGln, and 1 copy between tRNAGln and tRNAAla, along with the typical sets of genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) including regular tRNALeu(CUN) and the A+T-rich region (at least 923 bp). Only 1 copy of tRNALeu(CUN) differed by 1 bp from other 4 copies of tRNALeu(CUN). Each additional tRNALeu(CUN) is followed by nearly identical 68-bp long repeat sequence (95.6% identity). All 13 protein coding genes have typical start codons found in insect mitochondrial PCGs (2 ATA, 9 ATT, and 2 ATG).

Escape pattern of the ambrosia beetle Platypus koryoensis (Murayama) based on individual gallery was investigated. The survey was conducted in four oak forests located in Chuncheon, Hanam, Paju and Seongnam, and escape patterns from individual gallery were surveyed using emergence trap attached at the entrances of galleries formed in 2010. The number of adults caught in each trap was periodically counted from May 2011 to September 2012. The adults ambrosia beetle was caught over two years after infestation. The escape patterns in the first and second year after infestation showed different types; in the first year, escape of adults from gallery was observed one or two times whereas escape from gallery was observed two times in the second year with variation in aspect of escape density and timing. Moreover, variations in escape pattern between trees or between galleries in the same tree was observed. Colonization success rate in living tree was higher than that of dead tree, but the number of progeny per gallery in living tree was lower than that of dead tree. Colonization success rate in the galleries of second year was lower than that of first year, but the number of progeny per gallery of second year was higher than that of first year.

Mealworms, Tenebrio molitor (L.) is used as an important animal feed additive for growth promotion and health management, but potentially exposes to fungal infection. In this work, virulence of two species of entomopathogenic fungi against the insect, and the relationship between abiotic features and virulence were investigated. Secondly our consideration was also given to the effect of chemical fungicides on conidial germination for risk control. Between Beauveria bassiana (Bb) and Metarhizium roberstii (Mr) (previously M. anisopliae), Bb isolates had much higher virulence (~100% mortality in 3~4 days after the treatment), rather than Mr isolates in laboratory assays. Next, fungus-treated mealworms were kept at wheat bran at 20, 25, 30 and 35℃ with 3, 6, 9 times of water spray to the feeds for set-up of different humidity conditions. Inoculation of fungi to mealworms was conducted by fungal spray and feeding methods, which resulted in higher virulence in feeding method. In the feeding method, all temperature treatments except 35℃ showed high virulence against mealworms, but any significant relationship between virulence and humidity was not observed. In the chemical fungicide screening, fluazinam (CAS No. 79622-59-6) and mancozeb (8018-01-7) significantly inhibited the germination of Bb and Mr conidia. This work suggest that contamination of wheat bran with fungal pathogens, particularly B. bassiana may induce mycosis of mealworms, but introduction of effective fungicides possibly reduce fungal infection.

Worldwide studies on Apis cerana variation for biogeography and genetic diversity depended largely on a 86~93 bp-long mitochondrial non-coding region (internal spacer region) located between tRNALeu and COII (named as NC2), possibly due to higher variability among available markers. In order to incorporate the A. cerana occurring in South Korea into world extensive data, we also sequenced the NC2 from 118 A. cerana samples collected over nine Korean localities and 66 A. cerana samples over seven Asian localities, such as China, Vietnam, and Thailand. These data were combined with preexisting world data to scrutinize genetic relationships of A. cerana in South Korea to outside distributional range. Sequencing of 184 samples provided a total of ten haplotypes: five from Korea, six from China, one from Vietnam, and two from Thailand. Among them eight were new, whereas two were previously reported ones. Phylogenetic analysis of A. cerana NC2 haplotypes so far found including ours has confirmed the presence of four major groups of A. cerana (Asian mainland group, Sundaland group, Palawan group, and Luzon-Mindahnao group) and all haplotypes found in this study also were included in the Asian mainland group. In order to find further variable regions that can be used as sequence-based marker several mitochondrial non-coding regions and nuclear intron regions are in the middle of testing.

The aim of this experiment was to count excreted oocysts and to evaluate gross lesions and histopathological changes in broiler chicks infected with Eimeria; E. acervulina (Ea), E. maxima (Em), and E. tenella (Et). Twelve-day-old broiler chicks without infection with Eimeria spp. were used in this experiment. On day 12, the chicks in group Ea (n=3), Em (n=3), and Et (n=3) received oral inoculation with 5 × 10 sporulated oocysts of Ea, Em, and Et per 1 mL of tap water, while control chicks were injected with tap water of equal amount. The prepatent period (PP) for Ea, Em, and Et was four, five, and six days, respectively. To evaluate the progress of the coccidiosis, post-mortem analysis method and histopathological evaluation were performed two days after PP of Ea and Et, and three days after PP of Em at two-day intervals on one bird from each group. Intestinal segments (Ea, duodenum and upper intestine; Em, mid intestine; and, Et, cecum) of all chicks were obtained from the autopsied birds. Among pathological changes of coccidiosis, target parts of small intestine and caecal pouches showed mucosal petechiation and white plaques streaks in the duodenum. Histopathological evidence showed upper small intestinal villus preserved with different phases of parasite (zygotes, oocysts, macro- and micro-gametocytes) after infection with Ea in broiler chicks. Zygotes in the lamina propria and fusion of middle small intestinal villi in broilers infected with Em and invasion of the caecal crypt of oocysts and granulomatous inflammation of caecal submucosa in broilers infected with Et were found.

본 연구는 부존 조사료자원의 활용으로 수입 사료를 줄이고 사료비 절감에 기여하고자 갈대(Phragmites communis)를 공시하여 생육시기별사료가치와 재생 및 생산성을 구명하였다. 시험 1은 경기 안산 시화간척지 갈대 자생지에서2009년 5월부터 2010년 4월까지 7생육시기별사료가치를 분석하였으며, 시험 2는 천안 국립축산과학원 갈대 재배지에서 2011년 5월부터11월까지 10시기별로 수량과 사료가치 및 재생특성을 조사하였다. (시험 1) 5월부터 갈대의생육이 진행될수록 사료가치는 크게 낮아졌다.조단백질 함량은 2009년 5월, 7월, 9월, 2010년 4월 말에 수확하였을 때 각각 7.7%, 4.6%, 3.7%, 2.2%로, 상대사료가치 (RFV)는 각각 80.2,65.1, 61.8, 52.8로, 건물 소화율은 각각 59.3%,46.9%, 42.2%, 36.4%로 감소하였다. 반면 리그닌 함량은 7.1%, 9.9%, 12.0%, 13.2%로 크게증가하였다. (시험 2) 1차 수확한 갈대의 생육과 수량은 6월 하순에서 7월 중순 수확시 양호하였으며, 조단백질, RFV, 건물소화율 등 사료가치는 시험 1과 마찬가지로 생육이 진행될수록 낮아졌다. 1차 수확이 빠를수록 갈대의 재생은 양호한 경향이나 7월 말 이후는 차이가작았고, 8월 이후 재생은 매우 부진하였다. 연간 수량은 2011년 5월, 6월, 7월, 8월, 9월, 10월, 11월 말에 수확하였을 때 건물수량은 각각7,329 kg/ha, 12,527 kg, 9,593 kg, 8,279 kg, 7,649kg, 5,822 kg, 5,540 kg으로, 조단백질 수량은 각각 1,067 kg/ha, 1,201 kg, 1,046 kg, 533 kg, 377kg, 258 kg, 175 kg으로, 가소화 건물수량은 각각 3,924 kg/ha, 5,264 kg, 4,273 kg, 3,322 kg,3,352 kg, 2,195 kg 및 1,887 kg으로 낮아졌다(p<0.05). 본 연구에서 RFV를 기준한 갈대의조사료 (건초) 품질등급은 5월 갈대는 4등급 (다소 불량), 6~9월 갈대는 5등급 (불량), 10월, 11월 및 이듬 해 4월 갈대는 6등급 (부적합)이었으며, 재생 갈대는 모두 4등급 (다소 불량)으로분류되었다. 이상의 결과를 종합하여볼 때, 갈대는 늦어도 7월 안으로 수확․사료화 하는 것 이 양호한 가소화 건물수량과 볏짚 이상의 사료적 가치를 기대할 수 있어 권장되었다. 아울러 갈대의 재생은 전반적으로 불량하였고 1차수량이 연간 수량의 70% 이상을 점유하였으며,갈대는 연 1회 수확하여 사료화 이용이 바람직하였다.

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.

본 연구는 부존 조사료자원 이용으로 수입사료를 줄이고 사료비를 절감하고자 2010년도에 전남 해남(간척지), 강원 평창(강변), 원주(섬강변), 전남 영광(와탄강변) 등 4개 지역에서 갈대와 억새 등 야초 자생지를 탐색하고,수확 이용실태 조사와 함께 사료가치를 분석하였다. 전남 해남 해남목장 내 갈대(Phragmitescommunis) 위주의 야초 자생지는 300 ha로 예년에는 2회 곤포작업하여 연간 사일리지로2,300~2,500톤(ha당 8톤)을 생산하였으나 2010년도는 1회 곤포조제로 사일리지 2,000톤(ha당7톤)을 생산하였다. 유통가격은 롤 당 53천원으로 kg당 110원이었다. 곤포는 전량 자가소비하며, 외부 작업단에 롤 당 26천원의 작업비를지불하였다. 연간 조수입은 2억 2,260만원이며,소득은 1억원 수준(345천원/ha)이었다. 초장150 cm의 갈대는 조단백질 8.4%, 상대사료가치71.9 (건초 5등급), 건물 소화율 60.1%로 사료가치는 볏짚에 비해 약간 나은 것으로 평가되었다. 갈대 사일리지의 사료가치와 유기산 함량은 동일 지역에서 동일한 재료로 사일리지를조제하더라도 수확시기에 따라 차이가 컸으며(CP 4.7~6.4%, RFV 62.2~78.9, 건물 소화율41.9~53.9%), 갈대 사일리지의 품질은 수단그라스에 비해 불량하였다. 강원 평창 강변 자생갈대의 사료가치는 초장이 79, 117, 121, 142cm로 생육이 진행됨에 따라 낮아져, 조단백질은 각각 13.9%에서 7.6%로, 상대사료가치는90.7 (건초 3등급)에서 76.1 (건초 4등급)로, 건물 소화율은 72.9%에서 54.7%로 감소하였다.원주 섬강 주변 야초류는 곤포수확 작업에 어려움이 있었으며, 초장 130 cm의 갈대는 조단백질 8.5%, 상대사료가치 82.3 (건초 4등급), 건물 소화율 70.2%로 사료가치는 높았으며, 220cm의 성숙한 갈대는 조단백질 10.2%, 상대사료가치 65.1 (건초 5등급), 건물 소화율 48.9%로볏짚 대비 조금 나은 것으로 평가되었다. 전남영광지역에서는 와탄천 주변에 자생하는 갈대를 수거 이용하고자 시도하였으나 경제성이 없어 중단하였다. 이상의 결과를 종합하여볼 때,갈대, 억새 등 야초류는 현 상태에서 최대한수거 이용하되, 사료가치를 유지하고 가축 기호성이 양호한 시기에 수확하는 것이 중요하며, 최소한 볏짚과 비슷하거나 그 이상의 사료가치를 기대하기 위해서는 7월 중순 이전(늦어도 7월까지)에 수확 이용하는 것이 권장된다.

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.

The leaf beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) of the species collected from seven Korean localities. A total of 17 haplotypes (CACOI01 ~ CACOI17), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 16 sequence types (ITS2CA01 ~ ITS2CA16), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in COI gene sequence. Phylogenetically, the COI gene provided two haplotype groups with a high nodal support (≥ 87%), whereas ITS2 provided one sequence type group with a high nodal support (≥ 92%). The result of COI gene may suggest the presence of historical biogeographic barriers that bolster genetic subdivision in the species. Different grouping pattern between COI gene and ITS2 sequences were interpreted in terms of recent dispersal, reflected in the ITS2 sequence. Finally, finding of unique haplotypes and sequence types only from Beakryeng-Islet population was interpreted as an intact remnant of ancient polymorphism. As more samples are analyzed using further hyper-variable marker, further fruitful inference on the geographic contour of the species might be available.

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%).