When sperm penetrates into the ovum, hyaluronidase plays a role of hydrolyzing the hyaluronic acid present in the membrane surrounding the oocytes. The zona pelucida of the ovum is hydrolysed to facilitate sperm entry. Therefore, the aim of this study was to investigate the effects of hyaluronidase during the in vitro maturation in porcine oocytes. The cumulus-oocyte complexes (COCs) were cultured during in vitro maturation (IVM) medium containing 0 and 0.1mg/ml hyaluronidase for 44 h. Representative images of oocytes were captured after cultured for 0 h and 22 h by using a microscope. The area was quantified using a image J software. After 44 h of IVM, nuclear maturation stage was assessed by the aceto-orcein method. In results, cumulus cells expansion was no significant difference between control and hyaluronidase treatment groups in 0 h. However, after 22 h of IVM, in 0.1mg/ml hyaluronidase group, cumulus cells diffusion was significantly reduced than control group (p<0.05). After 22 h matured COCs, the cumulus cells were normally expanded in the control group, but there was a significantly lower 0.1mg/ml hyaluronidase group than control group (p<0.05). The nuclear maturation rate was treated with 0.1mg/ml hyaluronidase, it was significantly decrease than control group (p<0.05). In conclusion, our study indicated that hyaluronidase exposure could reduce nuclear maturation in vitro by reducing the expansion of cumulus cells. According to the results, we conjectured that hyaluronidase treatment disrupted the oocyte maturation by hydrolyzing the hyaluronic acid around the oocytes and it reduces the activity of the intercellular gap junction because it weakens cumulus cell bonds and interferes with communication. However, additional studies on hyaluronidase are needed. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Education) (2016R1D1A1B03931746).

In general, the shape of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage is important roles on meiotic maturation of porcine oocyte during in vitro maturation (IVM). Then, mitochondria produce reactive oxygen species (ROS) such as superoxide from electron transport system during oocyte maturation. ROS levels on oocytes are regulated by various antioxidant enzymes in cumulus cells (CCs). However, the effect of mitochondria derived superoxide production from CCs during IVM of porcine oocyte has not been reported. Firstly, we divided groups according to large number of CCs (Grade 1: G1) and small number of CCs (Grade 2: G2). Then, we counted cumulus cells of G1 and G2 oocyte by using haemocytometer. The oocyte maturation rate was significant decreased (p < 0.05) in G2 oocytes than that of G1 oocytes. We measured mitochondria derived superoxide in G1 and G2 COCs by using Mito-SOX staining. Mitochondrial superoxide was higher in G2 COCs than G1 COCs. Then, the mRNA expression levels of antioxidant enzymes (SOD1, SOD2 and PRDX3) in G2 COCs were decreased compared to G1 COCs. To reduce mitochondria derived superoxide, we used Mito-TEMPO as mitochondrial superoxide scavenger. Oocyte maturation rates in both G1 and G2 groups treated with Mito-TEMPO were increased than that of non-treated groups. Mitochondrial superoxide was lower in G1 and G2 groups treated with Mito-TEMPO than that of non-treatment groups. The mRNA expression levels of antioxidant enzymes in G1 and G2 COCs treated with Mito-TEMPO were increased compared to non-treated groups. Based on these findings, we suggest that reduction of mitochondria derived superoxide by Mito-TEMPO assists maturation competence in porcine oocytes.

Alpha-linolenic acid (ALA; n-3 18:3), a one of omega-3 fatty acid, is mainly contained in chloroplast of plant and ALA is an essential fatty acid, not synthesized in mammalian body, it must be supplied from foods. Polyspermy is especially high on in vitro fertilization (IVF) in pigs, which is a major obstacle to in vitro embryo production systems. In our previous study, when ALA was supplemented during in vitro maturation (IVM), the methaphase-II rate and gluthathione level was increased. The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) supplementation during IVM and subsequent of IVF in pigs. The cumulus-oocyte complexes (COCs) were submitted to IVM medium containing 0, 25, 50, and 100 μM ALA for 44 h. After 44 h of IVM, denuded oocytes were co-cultured with spermatozoa during 18 h. After 18 h of in vitro fertilization, oocyte were using aceto-orcein method, to evaluated penetration rate, monospermy (number of monospermy oocytes/total oocytes), and the IVF efficiency (number of monospermy/total penetrated oocytes). In results, 25 and 50 μM ALA groups were significantly increased on penetration rate compared with 100 μM ALA group (p<0.05). Similarly, monospermy rate were significantly increased 25 and 50 μM ALA groups than control group (p<0.05). IVF efficiency was no significant difference between control and ALA treatment groups. Our findings suggested that treatment of ALA supplementation during in vitro maturation (IVM) and subsequent of in vitro fertilization in pigs, ALA can increase IVF efficiency by effectively blocking polyspermy and increasing monospermy some mechanism in porcine oocytes. However, the study of mechanism by which ALA blocks polyspermy are needed, and this study suggests that ALA has a positive effect on in vitro production of porcine oocytes by decreasing polyspermy. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Education) (2016R1D1A1B03931746).

Korean horn beetles, Allomyrina dichotoma (Linnaeus, 1771) reared in many local farms are suffering from a fatal viral disease as reported in 2015, and recent sequencing analysis revealed that the virus is very closely related to Oryctes rhinoceros nudivirus. In a nationwide investigation, it was concluded that the virus can transmit vertically from an infected adult to the offspring by indirect manner, and 70 to 77% of young larvae dead in early stage were diagnosed as this virus positive. Here, we report for the establishment of on-site diagnosis method for the viral disease using loop-mediated isothermal amplification (LAMP) assay. A. dichotoma larvae were assayed for the OrNV infectivity with LAMP primers targeting OrNV_gp102 gene. To evaluate the LAMP specificity, two bacterial pathogens, Bacillus thuringiensis and Serratia marcescens, causing disease in A. dichotoma were tested along with OrNV. Only from the OrNV-infected larvae the reaction resulted positive. Also, to avoid DNA extraction process, the LAMP assay used diluted hemolymph directly and 50-fold dilution was set for diagnosis standardization and convenient on-site application for infected larvae from local farms. The LAMP diagnosis is fast and convenient for nontechnical user in the field and expected to stop this foreign virus spread all over the country.

Ethyl formate (EF) fumigation under green house condition is a new concept. Its concentrations inside and surrounding of the glass house (GH) and vinyl house (VH) were evaluated for 4 h (during daytime) and 12 h (during nighttime) after fumigation along with the evaluation of post-fumigation EF concentrations. The cumulative EF concentration × time (Ct) value of the 6 sampling positions in VH were 22.67 and 17.53 g·h/m3, respectively for day and night fumigation, which were 2.62 and 4.53 g·h/m3 respectively for day and night application in GH. The EF level (PPM) outside the VH and GH as well as after 20-min post fumigation were < 50 ppm revealing its safety level as its’ threshold limit value (TLV) is 100 ppm. The new technology using liquid EF fomulation could be a key option in smart-farm technologies in future. (Supported by PJ0133562018, RDA)

It is uncommon to fumigate the live plants in quarantine. Ethyl formate (EF) fumigation was carried out in a 0.275 m3 chamber in laboratory for 12 h (over-night) to evaluate the mortality of 4 insect pests and to evaluate the phyto-toxic effect on their host plants. The tested doses were 1, 1.5 and 2 mg/L EF with or without nitrogen gas (N2). The phytotoxicity was evaluated 7 days post-fumigation based on the damage index (DI) scaling 0-4. Mortality >80% were recorded against thrips, whitefly and aphid in all the EF or EF + N2 treatments. Two-spotted spider mite was highly tolerant (<20% mortality at the highest dose tested) to EF. Except for perilla and sesame (DI = 4), EF was non-toxic to other vegetable crops at 1.5 mg/L EF. However, 1.5 mg/L EF+N2 increased DI by 3 for tomato, watermelon and green pumpkin. This new method fumigating live plants certainly demands lots of further experiments to ensure its practical application in farm level. (Supported by PJ0133562018, RDA)

We investigated population densities of mosquitoes in Gyeongbuk region from March to November, 2017 and pathogens were detected from the mosquitoes. Mosquito populations were investigated at 3 sites in downtown and 1 site in cowshed in Andong, 3 sites in Yeongdeok downtown and 3 sites near migratory birds habitat in Sangju by collecting trap. Total collected female mosquitoes were 7 genus, 13 species, 9,892 individuals and relatively dominant ratios are Aedes vexans (40%), Ae. albopictus (20%), Culex pipiens (16%). Flavivirus were monitored in 9,125 collected mosquitoes by real-time PCR and no target flavivirus causing infectious diseases were detected in Gyeongbuk area in 2017.

Destruxins (Dtxs) are insecticidal cyclic hexadepsipeptides produced by the entomopathogenic fungus Metarhizium anisopliae. Media composition for dtxs production was optimized with industrial grade media. Glycerol and casein peptone were selected as a carbon source and a nitrogen source, respectively. Dtxs production varied with C/N ratios. High yields of dtxs were observed at C/N ratios ranging 0.3 to 1.5, with concentrations mostly higher than 800 mg/L. Low yields were caused by high C/N ratio ranging from 3.0 to 8.0, resulting in less than 500 mg/L. The highest yield of Dtxs was obtained with 2% glycerol and 3% casein peptone, showing 192.2 mg/L of dtx A, 911.1 mg/L of dtx B, and 113.3 mg/L of dtx E, respectively. These results indicated that dtxs production is highly influenced by C/N ratio, especially the content of nitrogen source.

Gonggeom-ji is designated wetland protected area by the Ministry of Environment of Korea in 2011 because of it’s high biodiversity and historic value. It contains a reservoir, paddy and forest site which has diverse niches for insects. Three transect lines of 50m were designated. Data were collected in May, October, November in 2017 represented as seasons for spring, summer and autumn respectively. Quantitative methods were conducted along each transect line by sweeping and pitfall trap. Based on this study total of 1079 individuals of insect fauna were collected, representing 170 species in 60 families and 8 orders. The highest diversity, richness and evenness index were observed in the forest site in May(4.77, 8.6 and 0.91 respectively) and the highest dominance index was observed in the forest site in November(0.64). The highest similarity index was observed in the reservoir site in May and August(0.519).

Heamaphysalis longicornis is a major vector for Severe Fever with Thrombocytopenia Syndrome (SFTS) virus and the density of the vector has been increasing because of the climate change. The incidence of fatalities due to SFTS is increasing every year. In this study, to evaluate the SFTS transmission by ticks, the density of ticks mediating SFTS was monitored. Tick was collected every month from four different sites (Grass land, Mountain path, Grave, Copse) in Andong with the traps containing dry ice as CO2 attractants. Among 2,572 ticks of 3 species; H. longicornis, H. flava, and Ixodes nipponensis were most abundantly collected from April to August. H. longicornis is the richest species (92.8%), whereas Ixodes nipponensis was the least species (0.8%). The 54.5% of the sample were nymph stage and female/male ratio was 64.3%. According to the pathogene analysis, SFTS virus was detected from H. longicornis adult and larvae stages collected in July and August. In July samples, SFTS virus was detected only from grassland site, but the virus was observed in the sample from all four sites in August. For effective prevention of SFTS, the tick density should be continuously monitored based on the onset time of SFTS with the consideration of habitate, habit & life history of ticks.

The mandible character of the Lucanidae is vary by considering of developmental difference in male individual specimen. Especially, species of Genus Cyclommatus Parry, 1862 presents strong mandibular allometry. In this research, The species group which related to Cyclommatus montanellus Möllenkamp, 1904 has been evaluated for the first time in this research with inner teeth composition and positioning to establish clear taxonomic key to each species.

상주는 귀농 도시이자 농업 중심 도시로써 농업기술원을 유치할 만큼 생태학적 연구가치가 있는 친환경 도시이다. 특히 경천섬은 낙동강 본류에 있는 상주보 인근에 위치하고 있어 낙동강 생태 환경 연구의 중요 지역 중 하나이다. 하지만, 아직 경천섬에 대한 생물다양성 연구가 부족한 상태이다. 섬과 산지의 생태환경이 다르기 때문에 생물의 다양성도 다를 것이라 간주하고, 경천섬 주변의 생태학적 가치를 재조명할 기초자료를 조사하고자 한다. 이에 경천섬과 인근에 위치한 청룡사 길목 산지에 서식하는 곤충 채집을 통해 생물다양성을 조사하여 비교분석하는 것을 목적으로 한다. 채집방법은 직접잡기, 쓸어잡기, 함정트랩, 흡충관을 이용하였으며, 함정트랩에 사용된 유인제는 소주1.8ℓ와 빙초산350㎖를 혼합하여 사용하였다. 2018년 4월~8월 중 3회 채집과정을 수행하여 목표개체수를 확보하였고, 채집된 곤충은 에탄올을 채운 conical tube에 넣어 냉동실에 보관하여 보존성을 높였다. 채집과정에서 잡은 곤충들을 건조시키고 유사종끼리 분류하고 동정하는 과정을 거쳐 다양도지수, 우점도지수, 종풍부도지수, 종균등도지수, 유사도지수를 계산할 것이다. 기존에 알려지지 않았던 경천섬 일대의 생물다양성을 세세하게 분석하는 과정을 통해 경천섬의 생태학적 가치를 드러내고, 경천섬 일대의 자연환경관리와 보존에 필요한 정보나 체계적인 관리체계를 구축할 기초자료를 제공하고자 한다.

본 논문에서는 한국어 쓰기 교육을 위한 교육용 게임으로서 text-to-scene 모델을 제안한다. 사용자 입력 문장에 대해 즉각적인 피드백을 제공하기 위해서, 제안하는 text-to-scene 모델은 사용자 입력 문장의 의미구조를 분석하여 장면을 자동으로 생성한다. 사용자가 문장을 교정하면서 자연스럽게 한국어 쓰기 능력이 향상될 수 있도록, 제안하는 text-to-scene 모델은 정답 장면과 사용자 입력 문장에 대한 장면을 함께 제공하므로, 사용자는 입력 문장의 어느 부분이 잘못되었는지 쉽게 파악하고 문장을 수정할 수 있다. 기존의 text-to-scene 연구에서 정교하게 장면을 생성하기 위해서는 필요한 자원 구축이 어렵고 처리 과정이 매우 복잡하다는 점을 고려하여, 제안하는 text-to-scene 모델은 외국인이 쉽게 이해할 수 있는 수준으로 장면을 단순하게 표현한다. 실험결과 격조사나 문장 요소 생략 등을 고려하여 15개의 단어를 임의로 조합하여 5개 어절 이내의 문장 1,048,576개 문장을 만들어 실험한 결과 제안하는 모델은 한 문장당 평균 14.94ms가 소요하면서 안정적으로 장면을 생성하였다. 외국인을 대상으로 한 작문 시험 결과, 종이를 이용한 작문 결과에서는 10개 문항 중 오답 문장이 평균 4.7개 나왔는데, 제안하는 text-to-scene 모델을 이용한 작문 결과에서는 한 문장 당 평균 1.56회의 미리보기를 하고 문장을 수정하여 0.87개가 나왔다.