West Nile Virus (WNV) is transmitted by infected mosquitoes. Vector mosquitoes usually acquire these pathogens fromfeeding on an infected host, and transmit the pathogens to a naive host during feeding events. To understand the virustransmission dynamics and to survey WNV throughout country, the present study has been conducted. We collected mosquitoesat urban parks in Seongnam, Wonju, Gunsan, Daegu, and Tongyeong using CDC light trap with Dry ice from April toSeptemper in 2017 (mosquito collecting is on going). Among collected mosquitoes, blood-fed mosquitoes were conductedblood meal identification assay and the other mosquitoes were subjected to virus detection using real-time PCR method.A total of 2,290 mosquitoes representing 6 genera and 15 species were collected. The most dominant species was Culexpipiens complex (42.1%) followed by Aedes albopictus (15.1%), Ae. vexans nipponii (14.6%), Ochlerotatus koreicus (9.8%),Cx. orientalis (6.5%), and Armigeres subalbatus (4.4%). The blood meal source were of mammal (93.3%), and birds (6.7%).So far, no WNV has been detected in any mosquitoes.

The members of the genus Flavivirus are noteworthy, as they cause infectious diseases in humans, such as Zika, denguefever, yellow fever, West Nile, and Japanese encephalitis. Due to the increased awareness of the public health risk posedby flavivirus-infected mosquitoes, mosquito collections were performed in six urban parks of South Korea, as the parksare designated for human recreation but also provide suitable habitats for mosquitoes. We examined the diversity andabundance of mosquito species and conducted molecular diagnostics for the detection of flavivirus infections. Monthlycollections were carried out in each park from March to August in 2017. A total of 4,851 mosquitoes (5 genera and13 species) were collected using BG-sentinel traps and then investigated for flavivirus infections. Pathogenic flavivirusinfections causing human diseases were not observed in the field-collected mosquitoes. However, insect-specific flavivirus(ISF) infections were detected in several mosquito pools. ISF has been previously known to enhance or suppress the replicationof medically important flaviviruses in co-infected mosquito cells. In this study, partial sequences of ISF were analyzed.However, further studies are needed in order to determine its genetic characterization and biological function in vivo.

Aedes albopictus is one species of mosquito transmitting flavivirus causing Dengue, Zika, and West Nile fever. Although it is an important disease vector, the genetic study of Ae. alpopictus populations has not been undertaken yet in South Korea. Here, we investigated the genetic variation of 99 Ae. albopictus individuals collected from 29 sites in nine provinces in 2016, through mitochondrial COI gene analysis. Haplotype analyses revealed seven haplogroups in South Korea. The main haplogroup, comprising 76 individuals (77.8%), was genetically identical to the one from Nagasaki. Two groups from Jeju Island (11) and the southern coast of South Korea (nine) were closely related to different Ae. albopictus strains from Kumamoto and Guangdong/Fujian, respectively. However, the others (four) were distinct from these two countries. No geographic divisions of populations were found in the study regions. The results suggest the possibility that the currently prevalent Ae. albopictus in South Korea, represents a part of the descendants that originated from nearby countries. However, more comprehensive investigations are needed to explain its movement routes.

Dengue is the most important arboviral disease in tropical and subtropical areas where 2.5 billion people are at risk of infection. Aedes albopictus will be a major vector transmitting dengue virus in Korea, where this virus overseas inflow is possible. We collected mosquitoes in Jeju, Busan, Gunsan, and Incheon using BG sentinel trap from April to October in 2016. Collected mosquitoes were conducted virus detection using real-time PCR method and analysis bloodmeal source of Ae. albopictus. A total of 15 species comprising 7 genera were identified and 4,854 female mosquitoes collected. The most dominant species ratio (SR) was 52.9% (Culex pippins complex) followed by 20.3% (Ae. albopictus), 10.8% (Ae. vexans nipponi) and Ochlerotatus dorsalis (3.8%). Dengue virus was not detected. Bloodmeal source of Ae. albopictus was mammals (80.9%) followed by birds (18.6) and amphibians (0.5).

We performed a survey for flavivirus infection and distribution of Aedes albopictus that known as Zika and Dengue virus vector using black–light trap and BG-sentinel trap around urban area in Korea. Mosquitoes were collected in 27 cities during March to November (twice a month) year 2016. Total numbers of mosquitoes collected 102,102 including 19 species 8 genera during collecting period. Total 21,467 Ae. albopictus was collected that 20,961(24.3%) by BG-sentinel trap and 506 (3.2%) by Black-light trap in urban area. Trap index(trap/night) of Ae. albopictus was showed highest in Hamyang (TI:992.3) and lowest in Taebaek (TI:0.3) there was only collected by Black-light trap. A total of 894 pools from all collecting Ae. albopictus were performed a Flavivirus detection. Flavivirus was not detected during study period. This study may provide basic information for surveillance of imported diseases (include Zika virus) and vectors in Korea.

West Nile Virus (WNV) is transmitted by infected mosquitoes. Vector mosquitoes usually acquire these pathogens from feeding on an infected host, and transmit the pathogens to a naive host during feeding events. To understand the virus transmission dynamics and to survey WNV throughout country, the present study has been conducted. We collected mosquitoes in Jeju, Busan, Gunsan, and Incheon using CDC light trap and BG Sentinel trap from April to October in 2016. Among collected mosquitoes, blood-fed mosquitoes were conducted blood meal identification assay and the other mosquitoes were subjected to virus detection using real-time PCR method. A total of 29,603 mosquitoes representing 8 genera and 19 species were collected. The most dominant species was Culex pippins complex (35.0%) followed by Cx. bitaeniorhynchus (12.2%), Armigeres subalbatus (11.2%), Aedes albopictus (10.8%), Ae. vexans nipponii (10.3%), and Ochlerotatus dorsalis (8.4%). The blood meal source were of mammal (70.4%), birds (29.0%) and amphibian (0.6%). WNV was not detected in any mosquitoes.

To identify whether higher expression of carboxylesterase (CbE) E4 in Myzus persicae is due to gene duplication, gene copy number was determined by quantitative real-time PCR. In addition, to determine the actual protein concentration of CbE E4 and it activity, Western blotting and activity staining were conducted. CbE gene copy number was highly correlated with carbamate resistance ratio (r2=0.934). However, CbE E4 expression level was little correlated with insecticide resistance ratio (r2<0.046) and no apparent correlation was observed among the gene copy number, protein quantity and total activity of CbE E4. Therefore, it was assumed that not only quantitative changing but also qualitative alteration of CbE E4 occurred in M. persicae. To investigate any potential alteration of CbE E4, mutation survey was conducted by sequencing of CbE E4 from various local strains of M. persicae. G137D and W251L mutations have been known as the main mutations associated with structural change leading to resistance. Interestingly, a new G134C mutation, which is in proximity of G137D mutation, was identified in the oxyanion hole of CbE E4. To predict the functional role of this mutation in resistance, 3-dimensional structure modeling was conducted. In summary, CbE E4 appears to be involved in resistance to both pyrethroids and carbamates as a nonspecific hydrolase or sequestration protein in M. persicae.

The green peach aphid (Myzus persicae) is a cosmopolitan pest of agricultural and horticultural crops and causes serious economic damages. M. persica has rapidly developed resistance to a wide variety of insecticides, including pyrethroids. Target site insensitivity mechanism mediated by two mutations (L1014F and M918T) on the para-type voltage-sensitive sodium channel (vssc) is mainly responsible for pyrethroid resistance. To predict the vssc resistance allele frequency, quantitative sequencing (QS) protocol was established. Frequency prediction equations generated from the plots of signal ratios and amplification critical time showed a high correlation coefficient (r2>0.993), indicating its high accuracy in prediction. QS results revealed that the kdr-type L1014F mutation is only present in Pyeongchang strain. No field strains of M. persicae possessed the super-kdr type M918T mutation. However, a novel M918L mutation was found by genotyping approach. The allele frequencies of M918L and L1014F were 0% to 53% in populations examined, and the level of M918L mutation frequency was closely related with pyrethroid resistance. Therefore, QS-based detection of M918L mutation frequency should faciltate the monitoring of pyrethroid resistance in the field.

The green peach aphid (Myzus persicae) is a serious pest of agricultural and horticultural crops all over the world. M. persica has rapidly developed resistance to a wide variety of insecticides, including carbamates. The E4/FE4 carboxylesterase is known to be involved in carbamate resistance. To compare the E4/FE4 carboxylesterase gene copy number, as a genetic resistance marker, between seven field strains, quantitative real-time PCR (qPCR) was performed. In addition, quantitative sequencing (QS) was employed to predict the frequencies of acetylcholinesterase (AChE) mutations (A301S and S431F) that are associated with target site insensitivity. All M. persica strains examined possessed the S431F mutation in the heterozygous state except for a susceptible strain, implying the possibility of AChE duplication. In contrast, no A301S mutation was found. Frequency prediction equation was generated from the plots of signal ratios and amplification critical time, which showed a high correlation (r2>0.996). QS analysis of M. persicae populations revealed that the allele frequency of S431F ranged 4% to 63%. Taken together, the AChE resistance allele frequencies determined by QS and the E4/FE4 gene copy number by qPCR should facilitate the detection and monitoring of carbamate resistance in M. persicae in the field.

Rhynchium brunneum is a widely distributed wasp species in South Eastern Asia. R. brunneum females were collected from rural provinces of Cambodia, and their total RNA and venom were extracted on site. To search for novel substances in venom, a subtracted cDNA library specific to the venom gland and sac was constructed. A total of 1118 expressed sequenced sequence tags (ESTs) were sequenced and assembled into 349 contigs (107 multiple sequences and 242 singletons). In this result, we found the putative neurotoxin (DTX protein precursor), antimicrobial peptides (teratocyte-specific caboxylesterase) together with typical major components of wasp venom (venom hyaluronidase, arginine kinase, phospholipase A2, serine/theonine protein phosphatase). Additional in-depth annotation would be required for further characterization of many unidentified genes found in the EST library.

Vespa tropica is a tropical species of Vespa found in Southeast Asia. V. tropica wasps were collected from rural provinces of Cambodia, and their total RNA and venom were extracted on site. To search for novel substances in venom, a subtracted cDNA library specific to the venom gland and sac was constructed and venom protein was analyzed by nano-LC-MS/MS. A total of 1127 expressed sequenced sequence tags (ESTs) were sequenced and assembled into 572 contigs (152 multiple sequences and 420 singletons). The short venom peptides were identified to be encoded from 5 contigs (43 ESTs) by proteomic analysis. In addition, putative antimicrobial peptides together with typical major components of wasp venom (venom allergen 5, mastoparan-like peptide, serine protease, and hyaluronidase) were identified in the EST Library. Additional in-depth annotation would be required for further characterization of many unidentified genes found in the EST library.

Insects or insect remains found in beer are one of major issues in consumer claim. Accurate estimation of inflow time isa critical factor for the settlement of such claims related with beer-contaminating insects but no reliable methods have been developed. In an attempt to establish a molecular marker-based diagnostic method, the degradation rates of 18S rRNA genes in the insectssoaked in 500 ml beer were investigated by quantitative real-time PCR (qPCR) over one month period at room temperature. Among the six insect species tested, the house fly (Musca domestica) and honey bee (Apis mellifera) revealed high correlations (r2=0.974-0.990) between the degradation of 18S rRNA gene and inflow time. In these insects, statistically significant distinction was possible between the samples stored in beer less than 14 days and more than 14 days. Other insects, including the fruit fly, common house mosquito, German cockroach and Indian meal moth, displayed poor correlations, which appeared attributed to the inefficient genomic DNA extraction likely due to small sample size or disintegration of body parts during storage in beer. With proper improvement in DNA extraction, this 18S rRNA-based diagnostic method would be applicable for estimating the inflow time of beer-contaminating insects.

피부는 스모그, 담배연기 및 UV와 같은 외부환경으로부터 신체를 보호하는 가장 큰 기관이며, 보호 기작 으로서 각질세포와 그 사이를 메우고 있는 세라마이드, 콜레스테롤, 지방산 등의 세포간지질이 라멜라 액정 구조 로 피부 장벽을 이루고 있다. 본 연구에서는 세포간지질 중 세라마이드 생합성과 관련되어 있는 세린-팔미토일 전이효소(serine-palmitoyltransferase, SPT) 발현을 western blot으로 확인한 결과, 제주산양산삼 추출물이 농도의존적으로 SPT 단백질 발현을 증가시킴을 확인하였다. 또한 제주산양산삼 추출물을 5% 함유한 제형을 2주간 피부에 도포 후 TEWL을 측정하였을 때, 제주산양산삼 추출물을 함유한 에멀젼 도포부위의 TEWL이 대 조군에 비해 유의적으로 감소하는 것을 확인하였다. 이 연구결과는 제주산양산삼 추출물이 SPT의 발현 증가를 통해 세포간 지질의 핵심성분인 세라마이드의 생합성을 증가시켰음을 보여준다. 따라서 제주산양산삼 추출물은 피부장벽기능을 개선시켜 TEWL 감소 효과를 나타내며, 이를 통해 화장품 분야에서 피부장벽 강화 및 보습소재 로서 사용될 수 있다고 사료된다.

Molecular markers are useful for selecting to include superior character genetic like as strong immune system and rapid growth in fish. The marker is also very important part of breeding technology in Olive flounder (Paralichthys olivaceus). Single nucleotide polymorphisms (SNPs) marker is already in use widely for genomic research and breeding. But this SNPs marker hardly has been validated for screening functional genes in Olive flounder. We study identify single nucleotide polymorphisms (SNPs) on Expressed sequence tag (EST) database, develop usable SNP marker and apply to wild sample and cultured of olive flounder. As a result, Out of total 4.327 ESTs, 693contigs and 514 SNP from total contigs were detected while these substitutions include 297 transitions and 217 transversions. 144 developed markers were applied in 16 samples (wild 8, culture 8), Out of total marker, only 32 markers had detected polymorphic in sample. Polymorphism of 32 markers was observed in the variety genes region involved in immunity and protein synthesis. And the 32 marker were identified 21 transitions, 11 transversions, and indel was not detected in polymorphic SNPs. The analysis on heterozygosity by sample showed 0.34 in wild sample and 0.29 in cultured sample. In conclusion, we was identified SNP and Polymorphism by designed new marker, it supports that development marker is suitable for SNP detection and diversity analysis in Olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

본 연구는 항균 활성을 지니는 식물추출물을 찾고 화장품에서 방부제로서 적용하기 위해 진행되었다. 디스크 확산법(disk diffusion method)을 통해 3가지 식물 추출물, 함박꽃나무(천녀목란, Magnolia sieboldii), 오배자(Rhus chinensis), 메타세콰이어(Metasequioa glyptostroboides)가 항균활성을 지니고 있음을 확인하였다. 세 물질의 최소저해농도 (MIC)를 측정한 결과 메타세콰이어는 0.3 ∼ 0.35 %, 함박꽃나무는 0.35 ∼ 0.4 % 농도에서 곰팡이의 생장을 억제하고 오배자는 0.45 ∼ 0.5 % 농도에서 세균의 생장을 억제함을 확인하였다. 또한 추출물 내의 항균 활성을 지니는 성분을 분리하여 분석한 결과 메타세콰이어에서 분리한 caryophyllene oxide와 caryophyllene, 함박꽃나무에서 분리한 costunolide와 dehydrocostus lactone, 오배자에서 분리한 ethyl gallate, ethyl-3-gallate 등이 항균활성을 지닌 물질임을 확인하였다. O/W 에멀션 제형에서 식물 추출물을 넣고 방부력을 확인한 결과 혼합사용 시 세균과 곰팡이 모두에 대한 방부효과가 있음을 확인하였다. 따라서 함박꽃나무, 오배자, 메타세콰이어 추출물의 혼합물은 기존의 화학 방부제를 대체할 천연 방부제로서 화장품에서 응용할 수 있을 것으로 기대된다.

ent-Kaurane- and ent-pimarane-type diterpenoids were isolated from the methanol extract of Siegesbeckia pubescens by column chromatography. Their structures were elucidated as ent-16α H,17-hydroxy-kau­ran-19-oic acid (1), ent-4,17-dihydroxy-16α -methyl-kau­ran-19-oic acid (2), ent-16β ,17-dihydroxy-kauran-19-oic acid (3), kirenol (4) and ent-16β ,17,18-trihydroxy-kauran­19-oic acid (5) by spectral analysis. The cytotoxicity of these compounds in Caki cells was assayed by a cell counting kit. Only one group treated with kirenol (4), an entpimarane-type diterpenoid, showed the inhibition of the cell growth in Caki cells.

This study is to identify the physiological traits of submergence-tolerant varieties of rice plants (Oryza sativa L.) in Yeongnam area, southeastern part of Korea, where the reduction of rice yield due to submergence is remarkably severe. In the present study, two tolerant varieties of rice plants were selected from over 30 rice varieties grown in under a 10-day period. The tolerant varieties selected from a submerged paddy field. As a control, one intolerant variety of rice plant was chosen. Of the tolerant variety Samgangbyeo, rather than Haepyungbyeo, had a lower dissolved oxygen consumption and maintained a higher dry weight than the intolerant variety. The leaf photosynthetic rates (LPS) of the two tolerant varieties were significantly higher than that of the intolerant-variety after four days of submergence treatment. These results indicate that lower dissolved oxygen consumption in a limited pool is prevented by ethylene formation in the tolerant varieties, which may be a mechanism of submergence tolerance.