The Ixodid ticks has been suspected to be the vector of severe fever with thrombocytopenia syndrome virus (SFTSV) in Korea. In order to confirm transovarial transmission of Ixodid ticks, a total of blood fed 284 ticks were collected from six host animals. They were identified as 3 genera and 4 species, Haemaphysalis longicornis (97.2%), H. flava (0.7%), Ixodes nipponensis (1.8%) and Amblyomma testudinarium (0.4%). Among them, 192 female adults hatched their eggs successfully. Our results showed a high prevalence of SFTSV among collected ticks (3.1%, 6/192 ticks), and we detected SFTSV from their larvae. This result suggests that the parent generation of H. longicornis can transmit SFTSV transovarially into their F1 progeny and H. longicornis might be one of major vectors of SFTSV in Korea.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne zoonosis and SFTS virus is transmitted to humans by tick bites. The aim of this study was to monitor tick population and its SFTS virus infection. During March 2014 to October 2015, ticks were monthly sampled using dry-ice bait trap and flag method from 4 collecting points in Boeun-gun. A total of 16,500 ticks, including 14,646 Haemaphysalis longicornis (88.7%), 1,825 H. flava (11.1%), and 29 Ixodes nipponensis (0.2%), were collected. Ticks were pooled (5,156 ticks in 456 pools) and tested by RT-PCR and nested PCR and the minimum infection rate of ticks was 0.27% (14 pools) consisting of 0.13% H. longicornis (6 pools), 1.19% H. flava (8 pools). This annual surveillance study will be needed for long-term monitoring of ixodid ticks in Korea.

Canine parvovirus (CPV), a member of the genus Parvovirus, family Parvoviridae, is a significant causative agent in Canine reproductive failure, causing serious economic losses in the pet industry. The major capsid protein, VP2 is the main target protein for neutralizing antibodies in CPV. When VP2 was expressed using baculovirus, it was produced abundantly and assembled into virus-like particles (VLPs) similar in size and morphology to the original virions. It was named as CPV-VLP. Additionally, p35 sequences of canine distemper virus (CDV) as T-helper epitope were fusion-expressed with each of N-term, C-term or both sides of CPV-VP2. Production of double antigenic recombinant protein and formation of VLPs were analyzed by SDS-PAGE and transmission electron microscopy, respectively.

Potato Virus Y (PVY) (Potyviridae: potyvirus) is one of the serious emerging virus of seed potato world-wide. It affects the seed potato by transmitting non-persistently via aphids. Here, we developed a simple PVY detection method which used the boiling technique for releasing of the viral RNA from aphid such as stylet and amplification by PVY specific primers located in the viral coat protein gene which suitable for various strains. This simplified method could save the time compared to earlier detection method due to the simplified RNA extraction step. Following this procedure, we tested this one step RT-PCR based PVY detection method by using three PVY vectoring aphid species (M. persicae, A. gossypii and M. euphorbiae) as well as other sucking type insect such as thrips (F. occidentalis). This PVY detection method is rapid, easy-to-use and suitable for large-scale testing in laboratories of seed potato.

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris, infected with an entomopathogenic fungus, Beauveria bassiana (Bb), and was further confirmed via the RACE method. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2 to 6 days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host.

West Nile Fever is one of most serious disease spreading throughout world since 1990. West Nile Virus (WNV) is transmitted by infected mosquitoes. The possibility of overseas inflow of this virus seems to be high in Korea. To know the WNV infection of mosquitoes, we collected mosquitoes in Jeju, Busan, Gunsan, and Incheon using CDC light trap and BG Sentinel trap from April to August in 2016. Collected mosquitoes were conducted virus detection using real-time PCR method. A total of 22,632 mosquitoes representing 9 genera and 18 species were collected. The most dominant species was Culex pippins complex (36%) followed by Aedes vexans (13%), Ae. albopictus (10%) and Ochlerotatus koreicus (10%). WNV was not detected in any mosquitoes until now.

Dengue is the most important arboviral human infection worldwide. Approximately 50-100 million cases occur annually and more than 2.5 billion people are at risk of infection. This virus is possibility of overseas inflow to be high in Korea. We collected mosquitoes in Jeju, Busan, Gunsan, and Incheon using BG Sentinel trap from April to August in 2016. Collected mosquitoes were conducted virus detection using real-time PCR method. A total of 15 species comprising 7 genera were identified and 3,696 female mosquitoes collected. The most dominant species ratio (SR) was 49.4% (Culex pippins complex) followed by 21.8% (Aedes albopictus), 13.6% (Ae. vexans) and Ochlerotatus dorsalis (4.8%). Dengue virus was not detected in any mosquitoes until now.

Ixodid ticks are notorious as the vector of severe fever with thrombocytopenia syndrome virus (SFTSV) in Korea. To confirm the transovarial transmission of SFTSV, we collected blood-fed adult ticks from animals in SFTS case reported area of Jeju island in 2015. A total of 476 ticks were collected from four host animal species, horse (81.3%), dog (9.0%), roe deer (7.1%) and wild boar (2.5%). They were identified as 1 genus and 2 species, Haemaphysalis longicornis (87.8%) and H. flava (12.2%). Among them, 131 adults hatched their eggs successfully. The body weight of females was highly correlated with the number of egg masses laid (P < 0.05). We confirmed that one adult H. flava was infected with SFTSV and its infection rate was 0.77%. The eggs of the SFTSV-infected adults also were infected with SFTSV and its transovarial transmission was confirmed. This result suggests that the parent generation of H. flava could transmit SFTSV transovarially into their F1 progeny and H. flava might be one of major vectors of SFTSV in Korea.

The Asian tiger mosquito, Aedes albopictus, is considered as potential vector of Zika virus in Republic of Korea (ROK). Vector control during mosquito season is one of critical factors for decline of viral transmission. Total 14 oversea travel-associated Zika cases by mosquito bite were reported throughout ROK from January to September 2016 and vector control and monitoring at surroundings of patient’s residences was carried out during three weeks after confirmation of the virus. Although population density rates of Ae. albopictus were remarkably various according to ecological surroundings, population density of Ae. albopictus near forest was higher than urban. All captured Aedes mosquitoes were used for detection of three flavivirus, Zika, Dengue and Chikunguya, using RT-PCR and any virus was not detected. Population density of Ae. albopictus decreased > 65% on average after vector control and in one area > 95% of population density decreased. Our data might reveal that vector monitoring and control at surroundings near residences of oversea travel-associated Zika patients might effectively prevent viral transmission by mosquito bite and naturalization of the virus in ROK.

The bluetongue is a serious vector-borne viral disease that affect wild and domestic ruminants. It is transmitted by midges belonging to the genus of Culicoides which count more than 1350 species. Since 1998, the disease has been spread in Europe and North Africa in 2000 including Algeria, while Korea reported serotype 1 of the virus on 2015. To know further about the existence and distribution of Culicoides species in Algeria, adult midges were collected from 17 different regions in Algeria from 2009 to 2015. The Interactive Identification Key of Culicoides (IIKC) has been used, followed by molecular identification by the sequencing of partial COI gene. At least 21 genetically different species have been identified. Among them, at least 5 potential virus vector species are present in Algeria. This study provides important insights for the understanding of genetic diversity of Culicoides and the existing potential of bluetongue spread in Algeria.

This research project provides the opportunity of foundation for Ministry of Agriculture y Ganaderia (MAG), University of El Savador (UES), and Centro Nacional de Tecnología Agropecuaria y Forestal (CENTA) and potato farmers of El Salvador to implement to achieve the project goal of poverty alleviation, nutrition improvement, and income increase of farmers through the establishment of sustainable self dependent national infrastructure for seed potato production and supply system and organized network of production of farmers. Kangwon National University has been designated by KOICA to implement the APP Track-II Project from April, 2015 to March, 2016. The support activities and research result include 'virus-free seed potato production facilities and laboratory equipment' provided to UES and CENTA, 'technology transfer of virus free seed potato production' such as making substrate, tissue culture, pathology test, subculture, mini tuber production, and some joint research result achievement, and various technological training conducted for participating researchers and local potato farmers for the virus-free seed potato production, table potato and seed potato cultivation technology, and the standard farming technology as well.

나리에 있어 Cucumber mosaic virus(CMV) 바이러스 저항 성 육종을 위해 주요 모본 및 부본으로 이용한 우수 유전자원 에 대해 바이러스 저항성 검정을 실시하였다. 바이러스 접종 원은 CMV-Taean으로 농업과학원에서 분양 받아 담배에 증식 하여 접종하였다. 바이러스 접종 식물체는 조직배양을 이용 하여 ‘Connecticut King’, ‘Party Diamond’, ‘Prato’, ‘Casa Blanca’, ‘Mona’, ‘Dublin’, ‘Rodrigo’, ‘Brunello’, ‘Avocado’ 품 종과 ‘FA08-17’ 계통에 대해 바이러스 무병주를 생산하여 실 험에 이용하였다. 바이러스 접종 효율을 높이기 위하여 접종 횟수를 1회와 2회로 나누어 실시하였으며 인산염 버퍼의 pH 를 6.9, 7.2, 7.5로 달리하여 접종하였다. 2회 접종, 인산염의 pH 6.9에서 접종 성공률이 높았다. ‘Casa Blanca’, ‘Avocado’ 품종의 잎에서 약한 모자이크와 노란색의 mottle 증상이 관측 되었으며 ‘Brunello’, ‘Rodrigo’, ‘FA08-17’에서 Hypersensitive response like 반응이 관찰되었다. ‘Brunello’, ‘Rodrigo’ 품종 에서 RT-PCR 검정 결과 바이러스가 검출되지 않았다. Reverse transcription-polymerase chain reaction(RT-PCR) 결과 바이 러스가 검출되었던 ‘Connecticut King’, ‘Prato’, ‘Casa Blanca’ 의 경우 Enzyme-linked immunosorbent assay(ELISA) 흡광도 가 낮게 나타났다. RT-PCR과 ELISA 검사 결과 잎과 구근에 서 CMV 검출이 다르게 나타남에 따라 나리에 있을 것으로 추측되며 바이러스 감염과 연관된 TOM1, OBE2, DND1 유 전자를 대상으로 발현 분석을 하였으나 유의한 결과를 얻지 못하였다.

Cordyceps species are known to produce numerous active components and are used for diverse medicinal purposes because of their varied physiological activities, including their anticancer, anti-inflammatory, and antimicrobial effects. Cordycepin, one of adenosine analogues, differs from adenosine in that its ribose lacks an oxygen atom at the 3' position. Cordycepin has been reported to be a main effector molecule in Cordyceps extracts that executes antiviral activities against several viruses including influenza virus, plant viruses, human immunodeficiency virus (HIV), murine leukemia virus, and Epstein-Barr virus (EBV). In addition, adenosine and its deaminase inhibitor showed strong antiviral potentials that were about 4,000 times more potent than the activity of the direct inhibitory effect by adenine arabinoside on Herpes simplex virus (HSV)-1, suggesting adenosine itself plays an important role to produce antiviral activities. In this study, we show that cordycepin and adenosine possess antiviral and antitumor activities against EBV and EBVassociated gastric carcinoma, respectively. Furthermore, we report epigenetic mechanisms used by cordycepin and adenosine to exert their antiviral and antitumor effects.

In this study, the virucidal efficacy of a fumigant (35% paraformaldehyde) against avian influenza virus (AIV) was examined. After AIV suspensions had been deposited on stainless steel carriers, the dried carriers were exposed to the fumigant in a 300-m3 test room for 3 h. Thereafter, all carriers were submerged in a neutralizing solution to scrape off the surviving viruses, and the respective suspensions were diluted. Each dilution factor was respectively inoculated into the allantoic membrane of five 10-day-old embryos. After incubation, AIV viability in the collected allantoic fluids was examined and the EID50 was calculated. The fumigant inactivated ≥5.7log10EID50 of AIV and was nontoxic to the embryos.

The most common method of vegetative propagation of virus free plantlets is the use of shoots meristem culture in solid medium culture. This study was to investigate the effect on liquid medium culture for the growth of virus-free sweetpotato plantlets. Single-nodes derived from meristem culture of sweetpotato was examined in this experiment and three sweetpotato varieties ‘Singeonmi’, ‘Sinhwangmi’, and ‘Sinjami’ was used. The growth of plnatlets was greater in liquid medium culture than that of solid medium culture after longer incubation in 3 varieties. The total fresh weight of 5 week old plantlets after planting in solid culture were 2.17 g (‘Singeonmi’), 2.49 g (‘Sinhwangmi’), and 2.18 g (‘Sinjami’), but the fresh weight in liquid medium culture was 3.87, 3.88, and 3.35 g, respectively. Leaf number of ‘Singeonmi’ and ‘Sinjami’ plantlets after 5 weeks of liquid medium culture was 21.1 and 22.6, respectively and liquid medium culture showed 3 and 6.2 more leaf number than that of solid medium culture. Plant height of ‘Sinhwangmi’ and ‘Sinjami’ plantlets after 3 weeks of liquid medium culture was 4.1 cm and 3.4 cm, respectively, and liquid medium culture showed 1.1 cm longer stem length than solid medium. Overall, liquid medium culture of sweetpotato plantlets was more effective than solid medium in terms of leaf and stem growth.

A novel insect-infecting positive sense single-stranded RNA virus, Riptortus pedestris virus-1 (RiPV-1), was found in the Riptortus pedestris transcriptome data by de novo assembly and further confirmed by RACE method. The genome of RiPV-1 consists of 10,554 nucleotides (nt) excluding the poly(A) tail and contains a single large open reading frame (ORF) of 10,371 nt encoding a 3,456 aa polyprotein and flanked by 71 and 112 nt 5' and 3' noncoding regions, respectively. RiPV-1 genome contains the consensus genome organization of picorna-like RNA helicase, cysteine protease, and RNA-dependent RNA polymerase (RdRp) array in that order from the 5' to the 3' end. From the phylogenetic analysis, RiPV-1 was clustered with unassigned insect RNA viruses, APV and KFV, which suggests that these three insect picorna-like viruses might constitute a novel group of insect-infecting RNA viruses. Tissue tropism analysis revealed that RiPV-1 was relatively abundant in the thorax, abdomen, midgut and fat body. Interestingly, RiPV-1 replication was enhanced by Beauveria bassiana JEF-007 infection that was quantified using qRT-PCR. This study identified a novel insect-infecting virus and provided further insight into the relationship between virus, fungus and host.

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.

Waterborne infectious disease is induced by several pathogenic microbes such as bacteria, viruses and protozoans, and the cases caused by viral infection is currently increasing. Water treatment process could reduce the number of virus in the water, but there were many difficulties to completely remove the virus particles from water. Therefore, the membrane separation technology which was reported to effectively remove pollutants from raw water has attracted increasing attention and demand. Since its efficiency has been introduced, demands for evaluation method toward the membrane filtration process are increasing. However, progression of the method development is slow due to the difficulties in cultivation of several waterborne viruses from animal models or cell culture system. To overcome the difficulties, we used adenovirus, one of the commonly isolated pathogenic waterborne viruses which can grow in cell culture system in vitro. The adenovirus used in this study was identified as human adenovirus C strain. The adenovirus was spiked in the raw water and passed through the microfiltration membrane produced by Econity, a Korean membrane company, and then the viral removal rate was evaluated by real-time PCR. In the results, the amount of virus in the filtered water was decreased approximately by 5 log scale. Because coagulant treatment has been known to reduce filtering function of the membrane by inducing fouling, we also investigated whether there was any interference of coagulant. In the results, we confirmed that coagulant treatment did not show significant interference on microfiltration membrane. In this study, we found that waterborne virus can be effectively removed by membrane filtration system. In particular, here we also suggest that real-time PCR method can rapidly, sensitively and quantitatively evaluate the removal rate of virus. These results may provide a standard method to qualifying membrane filtration processes.

애멸구(Laodelphax striatellus)는 rice stripe virus (RSV)의 매개충으로 벼에 큰 피해를 주는 해충이다. 본 연구에서는 애멸구의 장·단시형, 암·수, 약·성충에 대한 RSV 보독률과 이병률을 비교하였다. 애멸구의 장·단시형의 RSV 보독률은 각각 60.7%, 63.1%로 크게 차이는 없었다. 암·수에 대한 RSV 보독률은 각각 61.9%, 52.2%로 암컷의 보독률이 더 높았으나 유의성은 없었다. 약·성충의 보독률을 비교한 결과 각각 51.2%, 58.7%로 역시 크게 차이가 나지는 않았다. RSV에 감염된 애멸구에 노출된 건전한 벼의 이병률은 장시형은 53.3%, 단시형은 48.2%를 보였으며, 약·성충의 이병률은 각각 38.2%, 42.6%를 보여 유의성은 없었다. 반면 암컷은 50.5%의 이병률을 보이고 수컷은 22.3%의 이병률을 보여 암컷이 수컷에 비해 22.3% 이병률이 높아 유의성이 있었다. 또한 벼와 애멸구의 RSV 감염여부에 의한 애멸구의 발육기간은 건전한 벼에 RSV 감염 애멸구를 접종 했을 때 가장 긴 것으로 나타났으며, 건전한 벼에 건전한 애멸구를 접종 했을 때 발육기간이 가장 짧은 것으로 나타났다.