The purpose of this study was to determine the safe and effective method for preventing the occurrence of Oryctes rhinoceros Nudivirus in Allomyrina dichotoma on agricultural farms. There is a high demand for the use of A. dichotoma larvae in animal feed and as pet for educational purpose. Recently, we reported that OrNV is fatal virus diagnosed in A. dichotoma larvae in local farms in Korea. Mulberry leaves contain 1-deoxynojirimycin that represented anti-inflammatory, anti-virus, and anti-tumor effects. To prevent OrNV, we have fed the sawdust combined with 1% and 5% mulberry leaves powder to OrNV infected 2nd and 3rd stage larvae of A. dichotoma, and identified the mortality rate(%) during ten weeks. As a results, the 2nd stage larvae which were fed the sawdust combined with 5% mulberry leaves treatment recorded 60% mortality rate after ten weeks compared to the 100% mortality rate in the control. And the fatality rate of 3rd stage larvae which were fed 5% mulberry leaves treatment decreased 70% compared to the control. Therefore, application sawdust combined with mulberry leaves might be effective in the prevention and control of OrNV disease in A. dichotoma. Additionally, in the ten insect breading farms application test, OrNV virus disease have not appeared in the mulberry leaf powder treatment group.

Allomyrina dichotoma (order Coleoptera, family Scarabaeidae) is used for development of pharmaceuticals, pet or educational purposes and animal feedstuffs. The disease occurrence and distribution of Oryctes rhinoceros Nudivirus were investigated in Allomyrina dichotoma in Korea using PCR and analyzed the DNA seqeunces using BLAST(Basic Local Alignment Search Tool). The virus infected larvae were collected from 10 insect rearing farms in five different regions (Gyounggi, Chungbuk, Chungnam, Jeonnam, Daejeon). Frequency of OrNV virus infection appeared differently depending on the regions or rearing facilities (open field, vinyl house, indoor breeding system and etc.). The collected samples of Allomyrina dichotoma raised on open fields showed the highest possibilities of OrNV virus infection. The OrNV average infection rate of open fields rearing systems was 50.0%.

Mass breeding of Protaetia brevitarsis seluensis results in the entomopathogenic fungal infection, usually Metarhizium anisoplaie. A mixture of microorganisms (Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae) delayed fungal infection by M. anisopliae, which infected fewer P. b. seluensis when the microorganism mixture was added to sawdust as feed for P. b. seluensis for 30d, their mortality rate was approximately 35% less than that of the control group, which was fed sawdust without the EM. In addition, the growth of M. anisopliae on agar media spread with each bacterium as inhibited by up to 80%.

The purpose of this study was to identify the new fungal disease of Scopendra subspinipes mutilans (Chilopoda: Scolopendridae) from Jeju island in Korea. Scopendra subspinipes mutilans are also known the Chinese red head and average 20cm in length. It is used for traditional medicine for skin-disease and tumor and S. s. mutilans is recently raised in Jeju island to use healing properties. We isolated fungi from surface of infected S. s. mutilans and incubated in SDBA at 24℃ for 14days, and then identified the morphology of the fungi by light and electron microscopy. For the specific diagnosis of Metarhizium spp. in S. s. mutilans, 18srRNA of fungi was amplified by primers ITS4_MF and LR3_MR and sequenced by using the BLAST server at the National Center for Biotechnology Information. The result of the 18srRNA sequencing alignment was Metarhizium anisopliae (99.9%). The entomopathogenic fungi, M. anisopliae would be fatal cause of higher mortality to S. s. mutilans, and it is needed in vivo assay to confirm the pathogenic activities of M. anisopliae to S. s. mutilans.

The purpose of this stduy was to identify accurately and promptly the Microsporidia disease of bumble bees from Gangwon in Korea. Bumble bees are crucial pollinators of various crops and microsporidia are the critical infections of these hosts. Also, the various species of bumble bees have been used: Bombus terrestris, B. lucorum, B. occidentalis, B. ignites and B. impatiens etc. The symptoms of bumble bees infected microsporidia: their abdomens become distended, paralyzed and become sluggish and die early. We have identified the morphologies of the microsporidium by electron microscopy and found that the morphology of the microsporidium is rounded spore morph, with fairly small spores as described before in many other articles. For the specific and sensitive diagnosis of Nosema in bumble bees, we have developed the improved method of Polymerase Chain Reaction (PCR) conditions for expeditious diagnosis. Two pairs of primers were tested on N. bombi and the related microsporidia; Nosema apis and Nosema sp., both of which infect bumble bees and honey bees and further we have verified and analyzed DNA sequence data of N. bombi in bumble bees by using the BLAST server at the National Center for Biotechnology Information.

The purpose of this multiplex PCR assay is establishment and application for rapid and simultaneous detection of six pathogens related with insect diseases. Five pathogens were chosen based on the insect disease incidence rate in South Korea and specific primers of those pathogen were designed to detect insect diseases and test multiplex PCR for detecting Fungi; Beauveria bassiana(Bb), Metarhizium anisopliae(Ma), Bacteira; Bacillus thuringiensis(Bt), Pseudomonas aeruginosa(Pa), and Serratia marcescens(Sm). This research carried out the results detecting five kinds of insect pathogen of P. b. seulensis by multiplex PCR. Multiplex PCR is effective and save time to detect simultaneously these insect pathogens and multiple infections to prevent insect disease. In our study, using multiplex PCR, we demonstrated that P. b. seulensis was frequently infected with S. marcescens and co-infected with M. anisopliae in more than 80% of cases, indicating that such an analysis can be useful for pathogen identification, especially if different pathogens produce similar symptoms.