The α-Gal epitope (Galα1,3Galα1,4GlcNAc-R) is responsible for hyperacute rejection (HAR) during transgenic pig-to-non-human primate xenotransplantation. To overcome HAR after xenografts, it is essential for the inactivation of α1,3Galactosyltransferase (GT) gene by the homozygotic knocked out of GT-/- and the isoglobotrihexosylceramide synthase (iGb3s-/-). This study was performed to investigate the generation and characterization of the α1,3GT-MCP/-MCP+iGb3-/- transgenic cells. Ear fibroblast cells from the GT-MCP/-MCP pig were cultured and used to positive control. For iGb3s knock out, the Cas9-GFP-iGb3s vector was transfected into the GT-MCP/-MCP cells. The Cas9-GFP-iGb3s transfected cells were sorted and sequenced for the selection of GT-MCP/-MCP+ iGb3s-/- cells. Among the three sorted cell lines, one transgenic cell line was homozygously deleted 3 bases and 10 bases in each chromosome, respectively. To characterize an expression of α-Gal epitope, a wild type and the transgenic cells were measured by FACS Aria using BS-IB4 lectin antibody. The expression of α-Gal epitope in GT-MCP/-MCP cells (<0.01 %) were significantly down-regulated to the range of wild type (99.4 %) fibroblast cells (p<0.05). To analyze the function of iGb3s, α -Gal epitope expressions were measured for the GT-MCP/-MCP, GT-MCP/-MCP+iGb3s-/+, and GT-MCP/-MCP+iGb3s-/-. The range was 95.8%, 94.2%, and 75.8%, respectively. Interestingly, there was a negative range (16.2%) of α-Gal epitope -/- section in GT-MCP/-MCP+iGb3s-/-, compared to 2.74% of GT-MCP/-MCP+iGb3s-/+ and 1.4% of WT, respectively. Our results demonstrated that iGb3s-/-combined with GT-/- had a function to inhibit α-Gal epitope expression in pig cells. Further studies are needed to evaluate the functions of double gene knock out to minimize a HAR response after xenotransplantation.

The α-Gal epitope (Galα1,3Galα1,4GlcNAc-R) is responsible for hyperacute rejection (HAR) during transgenic pig-to-non-human primate xenotransplantation. There are genes related to the expression of α-Gal epitope such as α1,3Galactosyltransferase gene (GT-/-) and the isoglobotrihexosylceramide synthase (iGb3s-/-). This study was performed to investigate the expression of α-Gal epitope in the skin derived from GT-/- transgenic pig. The skin (7/1000 inches) was obtained by dermatome (Zimmer® Electric Dermatome) from one month old of wildtype (WT) and GT-/- piglets, respectively. The skins were fixed, dehydrated, cleaned, and embedded. To analyze the expression of α-Gal epitope, the paraffin section of WT and GT-/- were stained with BS-IB4 lectin and isoglobotrihexosylceramide synthase antibody. There was a strong BS-IB4 lectin signal in the skin of WT, but not detected in GT-/-. However, the iGb3s positive signals were stained in the skin of both WT and GT-/-. Taken together, it can be postulated that the knocked out of GT gene may not enough to inhibit the expression of α-Gal epitope. Further studies are needed to evaluate the functions of the double knock out of GT and iGb3s on the expression of α-Gal epitope.

For useful research animal to study human’s disease and for xenotransplantation donor, pig was studied to improve the quality of in vitro production (IVP). But, still the developmental ability of in vitro porcine embryos is still lower than in vivo embryos. Using a antioxidant is one of the strategy to overcome the drawback of in vitro producted embryos by protecting the oocyte from free radicals during in vitro maturation (IVM). Resveratrol, one of the plant-derived polyphenol antioxidants, have been used as effective antioxidants. Therefore, resveratrol treatment during IVM of porcine oocytes is expected to improve efficiency of the IVP by reducing free radical accumulation. In this study, we designed control (no treated) and resveratrol treatment groups (0, 2 and 4uM), evaluated maturation rate, cleavage rate, blastocyst formation rate and total cell number. Additionally GSH and ROS accumulation levels were measured via staining oocytes. In the results, maturation rate had not shown significant difference among the groups. However, in further development, not only the results of cleavage rate (0uM : 84.64±2.65 vs 4uM : 93.67±2.36, p<0.05) and blastocyst formation rate (0uM : 6.39± 0.90, vs 4uM : 13.67±2.32, P<0.05) were significantly increased in 4uM resveratrol treated group, and result of total cell number (0uM : 22.47±0.76 vs 2, 4uM : 30.35±1.76, 27.65±1.23, P<0.05) also shown significant difference in 2, 4uM resveratrol groups with control. GSH accumulated levels of matured oocytes in resvetrol treated groups were significantly higher than control. Meanwhile ROS levels of treated groups were significantly reduced [GSH (0uM : 142±10.49 vs 2, 4uM : 163.2±3.29, 169.7±0.94, P<0.05), ROS (0uM : 170.2±7.76 vs 2, 4uM : 118.6±7.90, 130±7.07, P<0.05)]. From these results, we conclude the treatment of resveratol improved further development of porcine embryos by regulating intracellular GSH, ROS levels during porcine IVM. Therefore, exogenous antioxidants such as a resveratol can be supportive substances for obtaining the improved quality of IVP.

Haemaphysalis longicornis is a major vector for Severe fever with thrombocytopenia syndrome (SFTS) throughout Republic of Korea (ROK). The ticks distribute at vast open fields and various kind of habitate. Control of the ticks with pesticides is almost impossible and personal protection such as use of commercial repellents is very critical for control. However occasional toxic effects of the commercial repellents have been reported that include central nervous system depression and urticaria. In our studies, repellency of three plant essential oils was assessed and was compared with two commercial repellents. Lavendual angustifolia demonstrated the highest repellency and higher repellency than DEET when was individually tested, but binary mixture of Cinnamomum zeylinicam and Juniperus verginiana showed significantly higher repellency with synergic effects than mixture of Lavendula angustifolia and Juniperus verginiana. Further study on repellency of the C. zelinicam in field need.

Previous laboratory repellent assessment with three plant essential oils against Haemaphysalis longicornis, a major vector for Severe fever with thrombocytopenia syndrome, revealed high repellency of Lavendual angustifolia and high synergic effects of Cinnamomum zeylinicam and Juniperus verginiana. Based on the previous laboratory repellent assessment field studies on repellency of the three plant oils were carried out with formulations such as cream type and spray type. 10% spray type of Cinnamomum zeylinicam demonstrated > 70% of repellency against H. longicornis. 10% spray type of Lavendual angustifolia showed > 80% of repellency. 10% spray type of binary mixture containing Cinnnamomum zeylinicam and Juniperus verginiana showed > 98% repellency against H. longicornis. Further studies on repellency of cream type of the plant oils might be carried out and compared with two commercial repellents.

Control of the vector mosquitoes is critical for implementation of patient’s decrease. Vector mosquito control has been mainly carried out using pesticides in Republic of Korea (ROK), which has developed the resistance against selected insecticides. Voltage-gated sodium channel (VGSC) and Acethylcholine esterase (AchE) are the target of several classes of insecticides. Genetic mutations of VGSC and AchE have been documented to confer resistances to pyrethroids and organophosphates in mosquitoes, respectively. In our studies, the insecticidal resistable level of An. sinensis from four malaria risk areas near Demilitarized Zone (DMZ) was measured. The partial sequence of An. sinensis voltage gated sodium channel (VGSC) IIS6 domain for Kdr gene and ace1 gene for AchE was sequenced. An. sinensis from two areas demonstrated significantly higher resistance to pyrethroids and mutation rates of Kdr gene were comparatively higher. In all tested areas An. sinensis showed high insecticidal resistance to organophosphates and > 70% of mutation rates of ace1 gene ware higher. Understanding the mechanisms of insecticide resistance among vector mosquitoes populations is very important to efficient and effective vector control.

Zika virus, mosquito-borne disease, caused by mosquitoes has been increased the importance. From March to September, twice a month from 7 different points (3 residential areas, 3 migratory bird sanctuary and 1 cattle shed) were collected using BG trap and BL trap. After identifying the mosquitoes collected, we confirmed the virus infection. Total 26,531 mosquitoes (6 genus 9 species) were collected, virus has been detected from the 3 species (Aedes vexans, Cuilex tritaeniorhynchus, and Amigeres subalbatus) of mosquitoes of them. It showed the highest peak in August, and then gradually decreased. The most common mosquito species was collected Aedes vexan (16,637) in the cattle shed.

Vespa velutina nigrithorax prey on various insect and especially hunts domestic honeybees, such as the European honeybee, Apis melifera. For that reason, V. velutina predation has a direct economic impact on apiculture. In particular V. velutina queens are able to establish of new colonies, so they are suitable for V. velutina management. This study was conducted to investigate the attractant of V. velutina Queen. Four traps were tested and combined attractant such as honeybee comb extraction, honeybee extraction, pollen, rice wine and sugar syrup for attraction efficacy. We was able to observe V. velutina Queen in late April to May. And Honeybee extraction, rice wine and sugar syrup baits (5 : 3 : 2) was the best combination for trapping queen on spring.

Bee venom is a complex mixture of toxic components that induces immediate local inflammatory and allergic responses. However, the presence and role of superoxide dismutase (SOD) in bee venom have not been previously investigated. Here, we provide the first demonstration that bee venom contains Cu,Zn SOD (SOD3), a novel extracellular component that promotes local inflammation. Bee venom SOD3 was shown to be an oxidant, rather than an antioxidant, that induces the inflammation-signaling molecule H2O2 in vivo. H2O2 plays a pathological role by triggering an immediate local inflammatory response. Furthermore, bee venom SOD3 significantly induced the activation of proinflammatory mediators (TNF-α and COX-2) and cytokines (IL-1β and IL-6) via the overproduction of H2O2 in mice. Our data demonstrate that bee venom SOD3 induced H2O2, which drives an immediate local inflammatory response, indicating a novel mechanism underlying bee venom-induced local inflammation.

We investigated pollination and foraging activities of honeybee (Apis mellifera L.) and bumblebee (Bombus terrestris L.) during flowering season of the asian pear (Pyrus pyrifolia N.) under different weather conditions. There was no significant statistical difference about the pollination activities of two species. However, the pollination activities of bumblebee were more active than those of honeybee under low temperature and rainfall period. The activities of honeybee and bumblebee were more influenced by temperature than other factors (i.e. illumination and wind velocity). Honeybee was more sensitive to temperature and illumination than bumblebee. At low temperatures (<20℃) on cloudy days (<30,000 lux) with a certain wind velocity (>4.0 m/s), the pollination activity of the honeybee was lower twice than that of bumblebee.

Pseudococcus longispinus, a notorious cosmopolitan pest species of mealybugs, known to be distributed indoors in Korea since 2002, is found to be mixed with another species, Pseudococcus orchidicola. Finding P. orchidicola as a pest of tropical plants in Korea is rather unexpected because of their main distribution in Pacific area and South Asia. However, all the available information from morphology, molecule and advices on identification from mealybug specialists indicated this is best matching P. orchidicola. Morphological, molecular and some biological notes on P. orchidicola are provided with some adult and nymphal images, and compared with P. longispinus. A full discussion mainly on identity and distribution of P. orchidicola is given.

Insect growth regulators (IGRs) are attractive pest control agents due to their high target specificity and relative safety to the environment. Recently, plants have been shown to synthesize IGRs that affect the insect juvenile hormone (JH) as a part of their defense mechanisms. We identified several JH agonists (JHAs) and antagonists (JHANs) from plant essential oil compounds using a yeast two-hybrid system transformed with the Aedes aegypti JH receptor as a reporter system. They showed high mosquitocidal activities with relatively low LC50 values and caused retardation of ovarian development in female mosquitoes. While the JHAs increased the expression of JH-induced gene, the JHANs caused reduction in the expression of the same gene. The compounds identified in this study could provide insights on the plant-insect interactions and may be useful for the development of novel IGR insecticides.

Bacillus thuringiensis (Bt) produces a variety of insecticidal crystal proteins and widely used as one of the most successful biological control agents. Recently, studies that introduce cry genes into crops to create pest resistance have made much progress, and the total area of land planted with Bt crops has increased substantially. In this study, pest resistance of 8 transgenic Bt rice events with a synthetic cry1Ac gene linked to rice rbcS-tp sequence were assessed under laboratory conditions. Bioassays were performed against Cnaphalocrocis medinalis, which is a significant pest of rice in Asia. C. medinalis larvae were shown to be susceptible to all eight events, even though there were differences between the causes of death. The results differed between developmental stages of the larvae, despite the fact that all 8 events led to high mortalities. These results may be a significant foundation for the evaluation of improved transgenic Bt rice.

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.

Beauveria bassiana JEF-007 with strong virulence against Riptortus pedestiris was selected for the Agrobacerium tumefaciens-mediated transformation(AtMT). AtMT generated two transformants, B1-06 and C1-49, showed significantly reduced virulence against R. pedestris. To identify the virulence-related genes, thermal asymmetric interlaced(TAIL) PCR and flanking region analysis were performed. From the analysis, two genes, Complex I intermediated-associated protein 30(CIA30) and Autophagy protein 22(Atg22), possibly related virulence in B. bassiana JEF-007. For the analysis of two putative virulence-related genes in JEF-007, hairpin RNA interference (hpRNAi) is under consideration. This work can provide the functional roles of the virulence-related genes in B. bassiana JEF-007.

Myeloid differentiation factor 88 (MyD88) is an intracellular adaptor protein involved in Toll pathway. In this study, we monitored the response of 4 key genes of the insect immune system against Beauveria bassiana JEF-007 in Tenebrio molitor using RT-PCR. To better understand the roles of Toll pathway in mealworm immune system, TmGPR and TmMyD88 was knocked down by RNAi silencing. Target gene expressions were decreased at 6 days post-dsRNA injection. Therefore, mealworms were compromised by B. bassiana JEF-007 at 6 days post-dsRNA injection. Silencing of the TmMyD88 and TmGPR resulted in reducing the resistance of the host to fungal infection. However, only dsTmMyD88 showed significant difference with dsEGFP by statistical analysis, which may be due to partial gene knock down of dsGPR. These results indicate that TmMyD88 is required in mealworms for survival against B. bassiana JEF-007.

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.