In perennial crops virus diseases are usually caused by mixed infections rather than by individual viruses. Understanding the contribution of each virus in disease development, the interactions between viruses and how each virus spreads in the field allows for development of control measures that are targeted for disease control rather than controlling all viruses in a complex. There are multiple types of virus-vector interactions and this information can be used to inform vector control strategies to manage virus diseases. Information on virus-vector interactions and insect biology for controlling a disease caused by a virus complex in raspberry will be presented. Understanding the biology of multiple vectors as well as multiple types of virus-vector interactions for a vector of multiple viruses will be presented as a model for managing virus disease in strawberry in different environments. The goal is to describe a systems approach for controlling virus diseases in vegetatively propagated crops from developing clean plants through to fruit production.

Human body and head lice are obligatory human ectoparasites. Although both body and head lice belong to a single species, Pediculus humanus, only body lice are known to be a vector of several bacterial diseases. The higher vector competence of body lice is assumed to be due to their weaker immune response than that of head lice. To test this hypothesis, immune reactions were compared between body and head lice following infections by two model bacteria, Staphylococcus aureus and Escherichia coli, and a human pathogen, Bartonella quintana. Following dermal or oral challenge, the number of these bacteria increased both in hemocoel and alimentary tract of body lice but not in head lice and the viability of the B. quintana was significantly higher in body louse feces, the major route of infection to human. In addition, body lice showed the lower basal/induced transcription level of major immune genes, cytotoxic reactive oxygen species and phagocytosis activity compared with head lice. These findings suggest that a reduced immune response may be responsible, in part, for the increased proliferation and excretion of viable bacteria which are associated with the high level of human infectivity seen in body versus head lice.

In the area of game graphics pipeline, efficiency of texture mapping process is very important. In this paper we proposed novel system of converting vector graphics data into shading language using the SVG vector graphics standard. Our system remove the dependance of special purpose authoring tool and utilize commercial vector authoring applications by using the XML-based standard. This study also can be used in the area of converting vector graphics from ‘implicit imaging model’ to ‘explicit imaging model’. We implemented this system in the RenderMan rendering architecture and we showed the result of applying vector graphics texture into 3D geometric model.

There are over 3,000 different species of mosquitoes throughout the world; currently 56 species are reported in Korea. Mosquitoes can act as vectors for many disease-causing viruses and parasites during blood sucking. Various species of mosquitoes are estimated to transmit various types of disease to more than 700 million people annually in Africa, South America, Central America, Mexico, Russia, and much of Asia, with millions of resultant deaths. At least two million people annually die of these diseases, and the morbidity rates are many times higher still. To prevent the spread of diseases, KNIH (Korea National Institute of Health) used three categorized methods; identifying or taxonomic analysis of mosquitoes, detecting virus caring mosquitoes, and detecting malaria from Anopheline mosquitoes. We have proved that taxonomic analysis using DNA barcording method (COI gene) is useful to complement identification of mosquito species. In detecting virus, we have reported Cx. orientalis as a new potential Japanese encephalitis virus (JEV) vector. We also reported that the Anopheline mosquito species composition and Plasmodium vivax infection rates in malaria hot spot in Korea.

현재 인간이 일반적으로 이용하는 의료용 단백질은 대개 동물세포 배양기술에 의하여 생산되고 있다. 그러나 제한된 특정 시설에서 생산되기 때문에 생산 단가가 높아서 전세계의 수요가 증가함에도 불구하고 일반화하는 데는 한계가 있는 실정이다. 이러한 문제를 극복하기 위하여 오래전부터 식물시스템을 이용하여 의료용 단백질을 생산하는 연구가 관심을 끌고 있다. 즉 쉽게 규모화할 수 있고 생산비용 효과도 경제적이고 동물세포보다 더 안전하게 의료용 단백질 생산할 수 있는 수단이 될 수 있기 때문이다. 이를 위하여 식물에 안정적으로 유전자를 핵 게놈과 엽록체 게놈에 형질전환 시키는 기술과 바이러스 운반체를 이용하여 일시적 발현을 유도함으로 의료용 단백질을 생산하는 기술이 개발되고 있다. 최근 해체 바이러스 기반의 운반체 개발은 재조합 단백질을 빠르고 일시적 발현으로 대량생산을 가능하게 하고 있다. 따라서 이 바이러스 발현시스템이 적절한 식물기반의 대량생산을 위한 플렛폼을 제공하고 있다. 따라서 본 총설은 식물로부터 의료용 단백질을 생산하는데 있어서 식물 바이러스발현 시스템 개발 및 이용에 대하여 기술하였다.

담배가루이(Bemisia tabaci)는 외래해충으로 바이러스벡터로 작용하여, 토마토의 토마토황하잎말림병바이러스(TYLCV)를 비롯한 약 100여종의 바이러스를 매개하는 중요한 해충이다. 본 연구에서는 VIGS vector를 이용하여 담배가루이 방제를 위한 target 유전자들을 선발하기 위 해 gateway system을 이용한 담배가루이 cDNA library 제작을 시도하였다. 첫 번째 방법으로 oligo d(T) primer를 사용하였을 때, 평균 약 1 kb의 insert와 1.4×10 4 cfu의 titer를 확인하였다. 그러나 insert size가 너무 커서 적절하지 않았다. 두 번째 방법으로 attB-N25 random primer를 이 용하고, sonication을 6초 실시하여 다시 진행하였다. 그러나 확인되는 insert size는 다소 컸고, 몇몇은 insert가 너무 작아서 밴드가 확인 되지 않았으며, 1.04×10 5 cfu의 titer를 확인할 수 있었다. 세 번째 방법으로는 oligo d(T) primer를 이용하였고, sonication을 2초 실시하였다. 그 결 과 300 bp~600 bp size의 insert가 확인되었으나, electro transformation을 사용한 첫번째, 두번째 방법에 비해 heat shock transformation을 사용하여 titer가 5.2×10 2 cfu로 매우 낮은 것을 확인 할 수 있었다. 결과적으로 cDNA library를 만들 때 먼저 random primer를 사용하여 First strand를 합성하여 poly A를 제거하고, 다음으로 sonication을 1초 실시하여 300~700 bp정도의 적절한 size의 insert를 생성하고, 마지막으로 electro-transformation을 실시하여 transformation 효율을 높인다면 VIGS vector에 적합한 cDNA library를 만들 수 있을 것으로 사료된다.

The sweetpotato whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is a disastrous pest in horticultural plants worldwide. B. tabaci is a species complex including at least 24 biotypes in the world. In Korea, B-biotype has been invaded in 1998, Q-biotype in 2005 and widely spread into the country. B. tabaci is also a vector of more than 100 plant viruses, especially begomoviruses. Since 2008, Tomato yellow leaf curl virus (TYLCV) has been invaded into Korea and severely damaged tomato cultivar in all over of the country. B. tabaci is the only vector insect of TYLCV. Here we demonstrated whether TYLCV influence on the vector physiology during virus transmission. Pesticide susceptibility of whiteflies on TYLCV acquisition was determined in B. tabaci using the two-layered parafilm feeding chamber which containing 20% sugar solution including different doses of imidacloprid. Our result showed that TYLCV-viruliferous whiteflies were more susceptible to imidacloprid ingestion than non-viruliferous whiteflies. This study suggests that plant virus can manipulate the physiological conditions of vector insects.

Brucellosis is an important and re-emerging zoonotic disease worldwide. The prevention of human infection is achieved predominantly through the control of brucellosis in agricultural animals, which in turn depends on accurate diagnosis and vaccination. However, conventional serological diagnosis of brucellosis has several limitations, and currently available vaccines for animals have several drawbacks, including the ability to cause infection in humans. Phosphoglycerate kinase (Pgk) is one of the specific proteins reactive with mouse sera in the early stage of Brucella infection, and deletion of the pgk gene in B. abortus strain 2308 resulted in extreme attenuation of this strain in vitro and in vivo. Furthermore, the B. abortus pgk mutant has been used as a live vaccine, and in challenge experiments, it induced protection that was superior to that conferred by commercial strains. In this study, the pgk gene from Brucella abortus 544 was successfully amplified and cloned into a maltose binding protein fusion protein expression vector (pMAL). The recombinant protein was expressed in Escherichia coli DH5α and purified. The immunogenicity of purified recombinant B. abortus 544 Pgk (rPgk) was evaluated by western blot analysis using Brucella-positive mouse sera. rPgk could be used as an antigenic component for future serological tests and potential vaccine development.

Many traditional fermented foods contain diverse bacteria and many lactic acid bacteria (LAB) play important roles for the fermentation of foods such as many dairy foods and kimchi. Especially, Leuconostoc mesenteroides is one of major bacteria in those fermented foods and the development of this species would be expected to be critical for strain improvement as well as the industrialization. Up to now, a lot of plasmids were isolated from Leuconostocs species including Leu. mesenteroides and a number of vector systems has been developed. Many plasmid vector systems using Leuconostocs spp. employ RCR (rolling circle replication) producing single-stranded DNA intermediates or sometimes takes theta replication. These plasmids include the sequences for Pre protein, recombination specific sites such as RSA (recombination site A) and RSB (recombination site B), and single strain origins for RCR replication. This information might be helpful to elucidate the improvement of Leuconostocs spp. and moreover the development of diverse products using Leuconostocs spp.

Leptotrombidium pallidum is the major vector mite for Orientia tsutsugamushi, the causative agent of scrub typhus, in Asian countries, including Korea. The genome size of L. pallidum was previously estimated to be 191 ± 7 Mb (Kim et al., 2014). Genomic DNA (gDNA) was extracted from a single female from a 9-generation inbred L. pallidum colony and used for whole genome amplification (WGA). The resulting amplified gDNA was used for the construction of paired-end and mate-pair libraries and sequenced using Illumina platforms (HiSeq2000 and MiSeq). An unamplified gDNA sample extracted from 20 female mites was also used for sequencing in parallel. More than 45Gb sequence reads from both paired-end and mate-pair libraries of the WGA gDNA were trimmed and then de novo assembled using the CLC Asembly Cell v.4.0 for contig assembly and SSPACE for scaffolding. The assembly generated approximately 6,545 scaffolds with N50 value of 92,945 and total size of ~193Mb, which was in a good agreement with our previous estimation. Repeat analysis showed that about 30% of genome (~58Mb) was masked as repeats, most of which were unclassified novel elements. For gene predictions, generated were the PASA models based on genomic alignments of RNA-seq reads from 4 different chigger mite samples (i.e. male, female, larva, and protonymph) and the GeneWise models based on genomic alignments of protein sequences from 4 closely related species with chigger mite. Independently, ab initio gene predictions were performed with AUGUSTUS and FgeneSH with custom trained matrices optimized for L. pallidum and GENEID with pre-trained matrix for Acyrthopsiphon pisum. By combining all together, 15,842 genes were predicted finally. Manual curation is in progress for various groups of genes, including chemosensory receptor genes, immune-related genes, acaricide target genes, etc.

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the most important citrus pest, because it serves as a vector of “Candidatus Liberibacter” species that cause huanglongbing disease. Thus, when exporting Rutaceae (citrus), exporting countries put them into quarantine. However, as the size of an imago is so small that only skilled experts can identify it using morphology-based species identification. A PCR-based assay was developed for monitoring psyllids using a rapid, using the DNA extraction from psyllid bodies and PCR amplification. Thus, this study aims to develop a DNA marker system with high species specificity and discrimination power that helps identifying psyllid. We analyzed the base sequence of mitocondria DNA. Based on this sequence, restriction sites were determined and a species particularity primer was made. Several polymorphic regions of mitochondrial DNA of both species were sequenced and used for developing specific restriction sites and polymerase chain reaction (PCR) primers. Also, species-specific PCR primers were devised to develop diagnostic PCR method for identifying the internal feeders. Base sequence results from PCR, which used psyllid species-specific primers, was analyzed by the Mealign program of the CLC Main Workbench analysis software, and used to develop a phylogenic tree. After analyzing this, Hemiptera proved to be allied species.

The genus Diadegma is a well known parasitoid group and some are known to have symbiotic virus, PDV. A novel IV was discovered from the calyx of D. fenestrale female. This virus was named as D. fenestrale Ichnovirus (DfIV). The encapsidated DfIV genome contains 65 circular DNA segments with an aggregate size of 247,191 bp. Based on BLAST analysis, a total of 120 ORFs were predicted as follows: rep; 48, cys-motif; 11, vinnexin; 10, vankyrin; 9, PRRP; 3 and other unassigned genes (39). These viral genes were expressed in lepidopteran hosts (Phthorimaea operculella and Plutella xylostella) after parasitization which means DfIV genome segments were integrated into lepidopteran hosts. This study was focused on this result. Based on gene expression profile, candidate promoter and integration motifs were selected and then, fused with eGFP as a reporter gene. Modified DfIV genome segment was ligated to a commercial containing f1 ori and Ampr gene to propagate in E. coli. We have named this fusion vector as pIN. The construction methodology of pIN and its application would be further discussed in detail.

Oak wilt disease caused by Raffaelea quercus-mongolicae was firstly reported at Sungnam city, Gyeonggi province, Korea, in 2004, and spread to several area of Korean peninsula. Raffaelea quercus-monglicae was transferred by ambrosia beetle, Platypus koryoensis, and caused oak wilt disease. Because oak wilt disease is mediated by ambrosia beetle, Platypus koryoensis, control of this disease concentrates on managing the ambrosia beetle population. Until now, several control methods such as felling and fumigation of dead oak trees with metham sodium, sticky-roll trap for catching emerged or attacking beetles, and oak tree logs for attraction of ambrosia beetle have been mainly used to control oak wilt disease in Korea. Although effective, current control methods have many side effects. Felling of oak tree is very difficult and dangerous to workers. To reduce the side effects of conventional control methods, new and safe types of control method for managing the ambrosia beetle populations are essential. In this study, we investigated the number of caught beetles, effect of height and direction of collection bottles by using newly developed mass capturing device for P. koryoensis to evaluate the effectiveness of this device as practical use in field.

묘목 생산성과 적절한 품질을 유지하기 위하여 시비는 매우 중요하지만, 수종에 따른 최적의 시비량을 결정하기는 어렵다. 이 연구에서는 경제림 육성 수종인 물푸레나무, 들메나무, 잣나무, 전나무를 대상으로 복합비료 1x(N 6.9 g m-2, P 3.1 g m-2, K 3.7 g m-2), 2x(1x의 2배), 4x(1x의 4배) 처리가 묘목의 생장과 양분변화에 미치는 영향을 양분벡터분석(Vector diagnosis)을 이용하여 정량화하였다. 시비량이 증가함에 따라 물푸레나무와 들메나무의 수고와 근원경은 증가하는 경향을 보였고 잣나무와 전나무는 시비량에 따른 차이를 보이지 않았다. 처리 1x, 2x, 4x는 대조에 비하여 물푸레나무의 건중량을 각각 43, 11, 87% 증가시켰고, 들메나무는 각각 36, 145, 143% 더 증가시켰다. 반면 잣나무와 전나무 건중량은 시비량에 따른 유의한 차이가 없었다. 시비에 대한 양분벡터반응은 수종과 시비량에 따라 상이한 경향을 보였는데, 물푸레나무 조직의 질소 농도는 생장 및 질소함량의 증가만큼 증가하지 않는 “양분결핍” 상태였고, 들메나무는 흡수한 것보다 생장이 증가하여 농도가 감소하는 “양분희석” 상태였으며, 잣나무와 전나무는 생장이 거의 증가하지 않으면서 농도는 약간 감소하는 “과량길항” 현상을 보였다. 본 연구는 물푸레나무와 들메나무는 처리 2x 또는 그 이상의 시비가 요구되고, 잣나무와 전나무의 경우 초기 시비를 하지 않아도 생장과 양분 흡수에 문제가 없음을 보여주고 있다.

Vector mosquitoes are related with various diseases on human, such as malaria (Anopheles sinensis (L.), An. sinensis), Japanese encephalitis (Culex tritaeniorhynchus (L.), Cx. tritaeniorhynchus), yellow fever (Aedes albopictus (L.)), dengue hemorrhagic fever (Ae. albopictus) and filariasis (Ae. albopictus), as well as nuisance insect pests (Culex pipiens pallens Foskal, Cx. p. pallens). Continued and repeated use of conventional insecticides such as organophosphorus and carbamates, pyrethroids has often resulted in the widespread development of resistance and has undesirable effects on non-target organisms and environment. Particularly, widespread insecticide resistance has been a major obstacle in the cost-effective integrated vector management program. In theRepublic of Korea (ROK), studies on mass vector mosquito control using insecticides were mainly carried out until 1970’s such as a fuselage-mounted ULV spray system to C-46 aircraft, effectiveness of ground aerosols ULV premiumsumihtion, a helicopter application of ULV Dibrom and residual toxicity of organophosphates in animal shelters for Japanese Encephalitis (JE) and Plasmodium vivax malaria vector mosquito control. Regional insecticide resistance of vector mosquitoes has been annually evaluated since ,early 1980’s, the first detailed study on susceptibility of seven mosquito species to 13 organophosphorus insecticides revealed the DDT and lindane resistance of Cx. tritaeniorhyunchus. Marked annual and regional variations of insecticide resistance have been observed. In comparative resistance, the resistance ratios of various insecticides were compared among each other using two strains of vector mosquitoes that were collected from same locality. An. sinensis from Koyang-si in 1992 and 1981 demonstrated >100 folds and >3000 folds of higher resistance to DDT and to bio-resmethrin. An. sinensis from Paju-si in 2008 demonstrated 30 folds of higher susceptibility to fenthion than those collected in seven years ago. Cx. tritaeniorhynchus from Gwanju in 2010 and in 1992 showed >100 folds of higher resistance to pyrethroids. In comparative regional resistance monitoring, Cx. p. pallens from six localities in the ROK in 2008 and six geospatially-distant field Cx. tritaeniorhynchus strains in 2011 showed marked regional resistance variations. Field populations of Cx. tritaeniorhynchus from agricultural areas demonstrated extremely higher insecticide resistance to pyrethroids than those from non-agricultural areas. Cx. tritaeniorhynchus populations from rice paddies had been under heavy selection pressure due to pyrethroids, which were used for a long time to control agricultural pests in the localities. These results indicate that careful selection and rotational use of these insecticides may results in continued satisfactory control against field populations of vector mosquitoes

Acquisition of plant viruses has various effects on physiological mechanisms in vector insects. Bemisia tabaci is the only known vector of Tomato yellow leaf curl virus (TYLCV), which is a serious virus affecting tomato cultivars. In this study, the lifespan of Q1 biotype was compared between non-viruliferous (NV) and TYLCV-viruliferous (V) whiteflies. Total lifespan from egg to adult death of NV whiteflies was 62.54 days but 10.64 days shorter in V whiteflies. We investigated the temperature susceptibility of B. tabaciby comparing mortalities as well as heat shock protein (hsp) mRNA levels between NV and V whiteflies. For this, NV and V whiteflies were exposed for either 1 or 3 h at 4, 25, and 35 °C. The mortality of V whiteflies was higher than NV ones following exposure at either 4 or 35 °C, but there was no significant difference at 25 °C. Analysis of the expression level of heat shock protein (hsp) genes using quantitative real-time PCR showed that both cold and heat shock treatments stimulated higher expression of hsps (hsp40, hsp70, and hsp90) at various rates in V whiteflies than NV ones, but there was no difference at 25 °C. All together, our results show that TYLCV acquisition accelerated the developmental rate and increased susceptibility to thermal stress in B. tabaci. Therefore, this modification may result in reduced vector longevity due to increased metabolic energy utilization. Our results provide insights into the complex interaction between vector fitness and thermal stress in relation to the acquisition and transmission of plant viruses.

Trombiculid mites are known to be the vector of tsutsugamushi disease by transmitting Orientia tsutsugamushi to human. Although the classification of trombiculid mites is necessary for vector surveillance, their classification by morphological observation is only possible at the larval stage and not easy because of similar shapes as well as tiny body sizes. Further the classification need the specimen production process, it takes much time and the accuracy of classification is changed according to the technology of the researcher. The internal transcribed spacers (ITS) regions of 8 trombiculid mite species were analyzed by amplification using tick common ITS primer sets. We designed molecular marker sets for the identification of five Leptotrombidium species, the lengths of marker L. orientale (1078 bp), L. pallidum (820 bp), L. palpale (1202 bp), L. scutellare (447 bp) and for L. zetum (621 bp) and three Neotrombicula species, the lengths of marker N. gardellai (264 bp), N. japonica (460 bp) and N. kwangneungensis (309 bp) based on alignment of ITS sequences. The markers will be helpful for exact classification of trombiculid mites. This study is the first report on molecular marker of ITS regions of trombiculid mites.

As more computer game integrates with video and working under mobile environments, it requires seamless bandwidth limited video transport environment including integrated game video compression and real time video conference between game users. Video transmission system consists of encoder and decoder. In communication environment, when the encoder transmits the stream data to the decoder, the bit-stream is alternated or lost because of various channel noises. Altered and lost bit-stream data cause the loss of the video data intensity value or motion vector (MV) value. These lost data cause serious damage of the video quality. In this paper, we propose MV recovery method because MV is essential information to keep the better video quality. To recover the lost MV, we use adjacent block’s MV as input data of Kalman filter to estimate the optimized value for current block’s lost MV. In our experimentation with the standard video image: Mobile&Calendar and Susie, our proposed method achieved 0.497dB ~ 1.898dB improvements on video quality over no MV recovery and achieved average 0.03dB ~ 0.725dB improvements over SRMC scheme.

Insecticide resistance and activation of the metabolic detoxification enzymes of female Culex pipiens pallens by the blood meal were assessed using a micro-application bioassay and micro-plate enzyme activity assays. Four group of Cx. pipiens pallens were used, a susceptible non-engorging group at seven days after emerging, SNE7 Cp; a resistant non-engorging group at seven days after emerging, RNE7 Cp; a resistant engorged group at one day after blood feeding and a resistant engorged group at seven days after blood feeding, REG7 Cp. Insecticide resistance of Cx. pipiens pallans was increased by the blood feeding. Based on LC50 values, SNE7 Cp demonstrated >50 fold of higher susceptibility to all tested insecticides when compared with RNE7 Cp. RNE7 Cp showed higher susceptibility to all tested insecticides than REG1 Cp and REG7 Cp with a relative susceptibility LC50 (SRLC50) of 25.8 to 50.0 and 25.0 to 48.8. In micro-plate enzyme assays, the metabolic detoxification enzyme activity of Cx. pipiens pallans adult females was increased by the blood feeding. Activation of non-specific esterases (EST), glutathione-S-transferase (GST) and mixed function oxidase (MFO) in RNE7 were higher than in SNE7 and the all tested enzymes in REG1 and REG7 demonstrated significantly higher enzyme activation than RNE7, except for activation of GST in REG1. Activation of MFO in REG1 and REG7 were 209.4- and 74.6- fold higher than in REG7, respectively. Non-specific esterases (EST) and glutathione-S-transferase exhibited < 10 fold of higher Rr values. These results may be significant in terms of the criteria that are used to evaluate resistance, because blood fed female mosquitoes may show enhanced expression of the resistance phenotype, possibly allowing for earlier detection of insecticide resistance.