본 연구는 주변 환경의 차이에 따른 화분매개곤충의 유입 특성을 파악하기 위하여 국립수목원 내 진화속을걷 는정원과 부추속전문전시원에 식재된 울릉산마늘의 화분매개곤충을 조사하였다. 2023년 5월 22일부터 6월 2일 까지 꽃이 70% 이상 개화하였을 때 포충망을 활용하여 8일간 곤충을 채집하였고, 각 전시원 별 식생(피도), 기후 (온도·습도·조도)를 조사하였다. 조사 결과 진화속을걷는정원에서 피도 60% 온도 26.4℃, 습도 31.5%, 조도 40953.6lx, 화분매개곤충 20과 450개체, 부추속전문전시원은 피도 90%, 온도 25.6℃, 습도 31.6%, 조도 6387lx, 화분매개곤충 15과 196개체로 나타났다. 온도와 조도가 상대적으로 높은 진화속을걷는정원이 채집된 곤충의 다양성과 방문 빈도가 높았다. 시간대별 곤충의 방문 빈도를 비교해본 결과 온도와 조도는 개체수가 증가할 때 같이 증가하는 경향을 보였으며, 습도는 반대의 경향을 보였다.

Republic of Korea (ROK) is operating the Integrated Environmental Radiation Monitoring Network (IERNet) in preparation for a radioactive emergency based on Article 105 of the Nuclear Safety Act (Monitoring of Nationwide Radioactive Environment). 215 radiation monitoring posts are monitoring a wide area, but their location is fixed, so they can’t cover areas where the post is not equipped around the Nuclear Power Plants (NPPs). For this, a mobile radiation monitoring system was developed using a drone or vehicle. However, there are disadvantages: it is performed only at a specific cycle, and an additional workforce is required. In this study, a radiation monitoring system using public transportation was developed to solve the above problems. Considering the range of dose rates from environmental radiation to high radiation doses in accidents, the detector was designed by combining NaI (TI) (in the low-dose area) and GM detector (in the high-dose area). Field test was conducted by installed on a city bus operated by Yeonggwang-gun to confirm the performance of the radiation monitoring system. As a result of the field test, it was confirmed that data is transmitted from the module to the server program in both directions. Based on this study, it will be possible to improve the radiation monitoring capability near nuclear facilities.

The red imported fire ant (Solenopsis invicta) is a species of ant native to South America. The fire ant was inadvertently introduced into USA, Australia, New Zealand, and other Asian countries including China and Taiwan. Since the first report of the fire ant in port city of Busan, Korea in 2017, it was found in many other cities of Korea in following year. To obtain the molecular information of this invasive species, total RNA was extracted from the abdominal segment of the ants collected in Incheon, and subjected to transcriptome sequencing. By using Illumina sequencer platform, 101 base pared-end sequencing generated 2 × 50,064,081 of raw reads to obtain 2 × 45.95 Gbase of quality filtered nucleotide sequences. The in silico cDNA library was constructed by Trinity de novo assembler followed by TransDecoder ORF finder and CD-HIT clustering program to streamline the library. The final version of cDNA library contains 20,442 contigs with protein coding capability. To survey the virome of this ant, these contigs were searched against the viral reference sequences from NCBI RefSeq database with BLASTN program. As a result, contigs which showed high sequence identities with several RNA viruses including previously reported SINV-2 were found from the fire ant. This virome information might give an idea of a shift of virological environment of this newly found ant isolate or population in Korea.

붉은불개미는 세계 100대 침입외래 해충 중의 하나로 우리나라에서는 관리 해충으로 1996년도에 지정되었다. 2017년 9월28일 부산항 감만 부두에서 국내 최초로 발견된 후, 2018년에 부산항, 평택항 과 인천항에서 발견이 되었고, 9월18일에 대구의 아파트 공사장에서 내륙에서는 처음으로 발견이 되었다. 대구에서 발견된 붉은불개미도 아파트 건설을 위하여 중국에서 수입한 석재에 혼합되어 수입된 토양과 식물체와 같이 혼입된 것으로 추측이 된다. 그러므로, 현재까지 국내에서 붉은불개미가 번식을 하여 토착화되었다는 증거는 없다. 현재까지 붉은불개미에 대한 종 판명은 전문가에 의한 형태적인 분류가 대부분이다. 그래서, 붉은불개미에 대한 확증을 위해서는 채집된 개체를 검역기관으로 이송하여 분자진단을 실시해야 하므로, 최소 이틀의 시간이 필요하다. 현재 항원항체 반응을 이용한 붉은불개미 진단 키트가 해외에서 개발되어 판매되고 있지만, 낮은 민감도로 최소 3~5개의 충체가 필요한 실정이다. 현재 판매되고 있는 진단키트가 가지는 문제점은 이를 대신할 새로운 키트의 개발이 필요함을 보여주고 있다. 신속한 붉은불개미의 검역현장에서 진단을 가능하게하기 위한 노력 중의 하나로 우리는 붉은불개미의 복부에서 발현되는 유전체에 대한 연구를 수행하였다. 본 발표에서는 그에 대한 현재까지의 연구 진행과 결과를 설명하도록 하겠다. (본 연구는 농림축산 검역본부의 연구비 지원으로 이루어졌음).

Nanopowders provide better details for micro features and surface finish in powder injection molding processes. However, the small size of such powders induces processing challenges, such as low solid loading, high feedstock viscosity, difficulty in debinding, and distinctive sintering behavior. Therefore, the optimization of process conditions for nanopowder injection molding is essential, and it should be carefully performed. In this study, the powder injection molding process for Fe nanopowder has been optimized. The feedstock has been formulated using commercially available Fe nanopowder and a wax-based binder system. The optimal solid loading has been determined from the critical solid loading, measured by a torque rheometer. The homogeneously mixed feedstock is injected as a cylindrical green body, and solvent and thermal debinding conditions are determined by observing the weight change of the sample. The influence of the sintering temperature and holding time on the density has also been investigated. Thereafter, the Vickers hardness and grain size of the sintered samples have been measured to optimize the sintering conditions.

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.

Euzophera batangensis (Lepidotera: Pyralidae) is seriously damaging trunks or branches of persimmon tree (Diospyros sp.). We tested the attractiveness of (Z)-9-tetradecen-1-ol (Z9-14OH) and (Z9,E12)-tetradeca- 9,12-dien-1-ol (Z9,E12-14OH) with single or blended baits at southern parts of Korea in 2014 and 2015. The monoene was not attractive at all at three places during the two years. In 2014, diene was equally or more attractive than the mixture in Jinju and Suncheon, respectively. In 2015 too, the attractiveness of diene and mixture was not different in Jinju and Munsan. Monitoring of the seasonal occurrence of E. batangesis with the sex pheromone components revealed that it occurred three times a year; the first occurrence from early April to mid May, the second from early Jun to mid July, and the third from late August to late September.

In the past four years, outbreaks of acute respiratory diseases associated with canine influenza H3N2 viruses in dogs and cats have been reported in South Korea and China. For prevention of disease from spread of the disease and for administration of timely medical treatments, including countermeasures for quarantine, use of a rapid and highly sensitive detection method are important to detection of the causative viruses. This study was conducted in order to develop a real time RT-PCR for the H3N2 subtype. It was based on primers targeting the highly homologous sequences of matrix, hemagglutinin, and neuraminidase genes. The detection limit of real time RT-PCR was 10 copies/ul with matrix and hemagglutinin genes, and 1 copy with neuraminidase genes, respectively. This real time RT-PCR was as sensitive as virus isolation in 52 clinical samples. The detection system developed in this study might provide more rapid and highly sensitive results than commercial rapid kits based on immunochromatographic assay.

Spider silks hold great potential as biomaterials with extraordinary properties. Here we report cloning and characterization of the major ampullate silk protein gene from the spider Araneus ventricosus. A cDNA coding for the partial major ampullate silk protein (AvMaSp) was cloned from A. ventricosus. Analysis of the cDNA sequence shows that AvMaSp consists of 240 amino acids of a repetitive region and 99 amino acids of a C-terminal non-repetitive domain. The peptide motifs found in spider major ampullate silk proteins, (A)n, (GA)n, and (GGX)n, were conserved in the repetitive region of AvMaSp. Phylogenetic analysis further confirmed that AvMaSp belongs to the spider major ampullate spidroin proteins. The AvMaSp-R cDNA, which contains sequences encoding for 240 amino acids of a repetitive domain, was expressed as a 22 kDa polypeptide of soluble form in baculovirus-infected insect cells. Recombinant AvMaSp-R was degraded abruptly by trypsin. However, AvMaSp-R was stable at 100 °C for at least 30 min. Additionally, the AvMaSp-R was stable at various pH values from 2 to 12 for at least 1 h. Taken together, our findings provide the molecular structure and biochemical property for A. ventricosus major ampullate silk protein as a biomaterial.

The silkworm-baculovirus expression system has distinct advantages, such as a high yield and safe usage in vertebrates. Here, we report a novel strategy for the large-scale production of a classical swine fever virus (CSFV) envelope glycoprotein E2 in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed native polyhedrin and approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra contained both the fusion protein and native polyhedrin were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Six-week-old female BALB/c mice that were immunized with the E2ΔC protein purified from solubilized recombinant polyhedraelicited CSFV E2 antibodies, which indicated that the CSFV E2ΔC protein from recombinant polyhedra was immunogenic. The virus neutralization test showed that the serum from mice that were treated with E2ΔC protein from recombinant polyhedra contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen.

Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.

Classical swine fever virus (CSFV) envelope glycoprotein E2 is the main target for inducing neutralizing antibodies and protective immunity in swine. Here, we report a novel strategy forthe large-scale production of a CSFV E2 subunit vaccine that demonstrates a high immunogenic capability in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (CSFV E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed an approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Mice that were immunized with the granule form of recombinant polyhedra or the soluble form of the fusion protein elicited CSFV E2 antibodies, which indicated that the recombinant polyhedra carrying CSFV E2ΔC were immunogenic. The virus neutralization test showed that the serum from mice that were treated with recombinant polyhedra or the soluble form of the fusion protein contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen and that the recombinant polyhedra containing CSFV E2ΔC as a granule antigen can be used as a potential subunit vaccine against CSFV.

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainlyinvolved in the xenobiotic metabolism and protection against oxidative damage. Most studies of GSTs in insects have been focused on their role in detoxifying exogenous compounds in particular insecticides. Here, we show the expression profiles of GSTs of the bumblebee Bombus ignitus in response to oxidative stress. We identified a sigma-class GST from B. ignitus (BiGSTS). The BiGSTSgene consists of 4 exons that encode 201 amino acids. Comparative analysis indicates that the predicted amino acid sequence of BiGSTS shares a high identity with the sigma-class GSTs of hymenopteran insects such as Apis mellifera (70% protein sequence identity) and Solenopsis invicta (59% protein sequence identity). Tissue distribution analyses showed the presence of BiGSTS in all tissues examined, including the fat body, midgut, muscle and epidermis. The oxidative stress responses analyzed by quantitative real-time PCR showed that under H2O2 overload, BiGSTS and BiGSTD (identified in our previous study) were upregulated in all tissues examined, including the fat body and midgut of B. ignitus worker bees. Under uniform conditions of H2O2 overload, the expression profile of GSTs and other antioxidant enzyme genes, such as phospholipid-hydroperoxide glutathione peroxidase (Bi-PHGPx) and peroxiredoxins (BiPrx1 and BiTPx1), showed that other antioxidant enzyme genes are acutely induced at 3 h after H2O2 exposure, whereas BiGSTS and BiGSTD are highly induced at 9 h after H2O2 exposure in the fat body of B. ignitus worker bees. These findings indicate that GSTs and other antioxidant enzyme genes in B. ignitusare differentially expressed in response to oxidative stress. Taken together, our findings indicate that BiGSTS and BiGSTD are oxidative stress-inducible antioxidant enzymes that may play a role in oxidative stress response.

Peptidoglycan recognition proteins (PGRPs) are pattern recognition molecules of the innate immune system that recognize peptidoglycan, a unique cell wall component of bacteria. Here we cloned and characterized PGRP-S from the bumblebee Bombus ignitus (BiPGRP-S). The BiPGRP-S gene consists of four exons encoding 194 amino acid residues. Comparative analysis indicates that the predicted amino acid sequence of BiPGRP-S shares high identity with enzymatically active PGRP-S proteins and contains the amino acids required for amidase activity. BiPGRP-S in B. ignitus worker bees is constitutively expressed in boththe fat body and epidermis, and it is secreted into the hemolymph. Quantitative real-time PCR assays revealed that in both the fat body and epidermis, the BiPGRP-S gene is highly induced by an injection of Bacillus thuringiensis. In addition, recombinant BiPGRP-S expressed as a 19-kDa protein in baculovirus-infected insect cells can bind to B. megaterium and B. thuringiensis but not to Staphylococcus aureus, Escherichia coli or Beauveria bassiana. Consistent with these data, BiPGRP-S shows antibacterial activity against B. megaterium and B. thuringiensis. These results indicate that BiPGRP-S is an inducible protein that may be involved in the immune response against bacterial infection of the genus Bacillus as an amidase-type PGRP-S.

Phospholipid-hydroperoxide glutathione peroxide (PHGPx) is an antioxidant enzyme that reduces lipid hydroperoxides in biomembranes. Here, we cloned and characterized cys-PHGPx from the bumblebee Bombus ignitus (Bi-PHGPx). The Bi-PHGPx gene consists of 4 exons, encoding 168 amino acid residues with a canonical cys-codon at residue 45 and active site residues Gln82 and Trp134. Recombinant Bi-PHGPx, expressed as a 19 kDa protein in baculovirus-infected insect cells, exhibited enzymatic activity against PLPC-OOH and H2O2 using glutathione as an electron donor. Tissue distribution analyses showed the presence of Bi-PHGPx in all tissues examined. Bi-PHGPx transcripts were upregulated by stresses, such as wounding, H2O2 exposure, external temperature shock, and starvation. Under H2O2 overload, the RNA interference (RNAi)-induced thioredoxin peroxidase (BiTPx1)-knock-down B. ignitus worker bees showed upregulated expression of Bi-PHGPx in the fat body. These results indicate that Bi-PHGPx is a stress-inducible antioxidant enzyme that acts on phospholipid hydroperoxide and H2O2.

Bee venom contains a variety of toxic enzymes and peptides. One of the major components of bumblebee venom is bombolitin, which is the most abundant venom constituent and biologically similar to melittin. Here, we first show the molecular cloning and antimicrobial activity of the venom bombolitin from the bumblebee Bombus ignitus. The B. ignitus venom bombolitin gene consists of 2 exons, encoding 56 amino acid residues. The bombolitin purified from B. ignitus venom was the 2104 Da mature peptide with 18 amino acid residues, which are created by cleavage of the probombolitin domain between Ala38 and Leu39. We examined the pattern of bombolitin expression to confirm that it is a component of bumblebee venom. B. igniutus venom bombolitin exhibits venom gland-specific expression. We also investigated the venom bombolitin for antimicrobial properties against bacteria and fungi. The venom bombolitin showed high antibacterial activity against both Gram-negative and Gram-positive bacteria. Most interestingly, the venom bombolitin showed high antifungal activity against Fulvia falva, a leaf mold, and Alternaria radicia, a black rot. These antimicrobial profiles of B. ignitus venom bombolitin reported herein will be useful in the application for potential antimicrobial agents.

We cloned and characterized two peroxiredoxins (Prxs), BiPrx1 (a 1-Cys Prx) and BiTPx1 (a 2-Cys Prx) from the bumblebee Bombus ignitus. The BiPrx1 gene consists of 5 exons, encoding 220 amino acid residues with one conserved cysteine residue. The BiTPx1 gene consists of three exons, encoding 195 amino acid residues with 2 conserved cysteine residues. Recombinant BiPrx1 (27 kDa) and BiTPx1 (25 kDa), expressed in baculovirus-infected insect Sf9 cells, reduced H2O2 in the presence of electrons donated by dithiothreitol. Unlike BiTPx1, however, BiPrx1 did not show reduction activity when thioredoxin was used as the electron donor. Both BiPrx1 and BiTPx1 protected super-coiled DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Tissue distribution analyses showed the presence of BiPrx1 and BiTPx1 in the fat body, midgut, muscle and epidermis, but not in the hemolymph, suggesting that BiPrx1 and BiTPx1 are not secretable. When H2O2 was injected into B. ignitus bees, BiPrx1 and BiTPx1 transcripts were acutely up-regulated in the fat body tissues. We also demonstrated regulation of BiPrx1 and BiTPx1 expression via reduction of transcript levels in the fat body with RNA interference (RNAi). Under H2O2 overload, the RNAi-induced BiPrx1 knock-down B. ignitus worker bees showed up-regulated expression of BiTPx1. Reciprocally, BiTPx1 RNAi knockdowns showed up-regulated BiPrx1 expression in the fat body. These results indicate that loss of expression of BiPrx1 or BiTPx1 is compensated by up-regulation of expression of the other peroxidase in response to H2O2 overload.

Monochamus alternatus and M. saltuarius were reported as the vectors of Bursaphelenchus xylophilus, pine wood nematode in Korea. According to Kwon et al. (2006), each of 2 species has occupied their own regional distribution : M. saltuarius in southern part including Jeju island and M. alternatus in mid-northern part of Korean peninsula. We measured the supercooling point (SCP) of 2 species (laboratory-reared populations) by each of developmental stages. The SCPs of 2nd, 3rd and 5th instar larvae of M. saltuarius were -7.68±0.19℃, -7.02±0.69℃, -4.93±1.34℃ each of stages. On the other hand, the SCPs of 3rd, 4th, 5th instar larvae and pupae of M. alternatus. were -4.46±1.12℃, -5.94±1.33℃, -7.83±1.44℃, -9.53±1.78℃ each of stages. The SCPs of M. saltuarius larvae generally was lower than that of M. alternatus. The pupae of M. alternatus and 2nd instar larvae of M. saltuarius had the lowest SCP among measured samples. On the other hand, the highest SCP were recorded in 2nd and 5th instar larvae, each. This result shows that regional distribution of 2 beetles may be associated with the adaptation capacity to low temperature represented by the SCP as well as the developmental temperature. However, beetles experimented were not collected from pine forest fields. In further study, we are planning experiments with field populations and all developmental stages.

핵연료 운반용 실린더의 재사용을 위한 용기세척공장의 제염공정에 대한 성능을 평가하기 위하여 Na2CO3 + H2O2 혼합용액의 조합을 약간 달리한 2회의 시험을 실행하였다. 각 시험은 모두 일련의 5 단계에 걸쳐 실시되었다. 우라늄 제염의 주 화학종은 Na4UO2(CO3)3 로 식별되었다. 그리고 첫 단계에서의 세척액은 물이었으며, 이 단계에서 50% 이상의 우라늄이 제염되었다. 그 이후로는 단계가 더해 갈수록 우라늄의 제염양은 지수함수적으로 감소하는 경향을 나타내었으며, 화학양론적으로 제거된 우라늄에 비하여 투여된 Na2CO3 의 양은 과다함을 나타내었다. 이러한 결과들에 의하면, 공정최적화를 통하여 Na2CO3 의 투여량 감축, 세척폐액의 감량, 제염단계 축소 등을 꾀할 수 있을 것으로 판단된다.