Earthworms have been employed in traditional oriental medicine for the treatment of neurological disorders, as anticonvulsants, analgesics, and sedatives. In fact, earthworms are currently used as a medicinal agent in China, South Korea, Japan, Taiwan and North Korea. However, both the species and the genus of the earthworms registered in the pharmacopoeia of each country are different. Neural injury is induced by oxidative stress, inflammation, and apoptosis. The role of various synthetic chemicals of earthworms as antioxidant and anti-inflammatory agents have been studied and earthworm extract and its components have been shown to protect nerve cells and restore nerve function in various preclinical neuronal damage models. We employed earthworm extracts to provide prevention and treatment strategies for many neurodegenerative disorders including Parkinson's disease, mild cognitive impairment, cerebral infarction, and peripheral nerve damage. In this study, we investigated the effects of earthworm extracts and its components to explore their prophylactic and therapeutic effects in various neuropathic models. We used earthworm resources to provide prevention and treatment strategies for many neurodegenerative disorders including Parkinson's disease, mild cognitive impairment, cerebral infarction, and peripheral nerve damage. We summarized the protective effects of both earthworms and their extracts on neuropathies. The current study identified some earthworm components to be used in treatment and prevention strategies for nerve disorders and could be helpful for the development of new therapies for intractable diseases.

Roller Compacted Concrete Pavement (RCCP) is placed by roller compaction of a mixture of less cement and unit water content and more aggregates and provides excellent early strength development with the help of interlocking of aggregates and hydration. The unit cement content of RCC pavements accounts for 85% of conventional pavements, with low drying shrinkage. As low drying shrinkage leads to smaller crack widths than ordinary concrete, RCC pavements can help elevate reflecting crack resistance if applied to a base layer of a composite pavement system. In a composite pavement with an asphalt surface laid over a concrete base, pavement temperature change is important in predicting pavement performance. As movement of the lower concrete layer is determined by temperature depending on pavement depth, temperature data of the pavement structure serves as an important parameter to prevent and control reflecting crack. Among the causes of reflecting crack, horizontal behavior of the lower concrete layer and curling-caused vertical behavior of joints/cracks are considered closely related to temperature change characteristics of the lower concrete course (Baek, 2010). Previous studies at home and abroad about reflecting crack have focused on pavement behavior depending on daily and yearly in-service temperature changes of a composite pavement (Manuel, 2005). Until now, however, studies have not been conducted on initial temperature characteristics of concrete in composite pavements where asphalt surface is placed over an RCC base. Annual temperature changes of in-service concrete pavements go up to 60 ℃, and those of asphalt overlays become around the twice at 110 ℃. This study evaluated initial crack behavior of composite pavement by investigating pavement temperature by depth of an RCC base and analyzing joint movement depending on change to temperatures of continuously jointed pavements. Findings from the study suggest that in composite pavements and asphalt overlays, time of laying asphalt has an important impact on crack behavior and reflecting crack.

Beauveria bassiana (Bb) is a potential biopesticide against western flower thrips. To understand molecular characteristics of two Bb isolates (ERL836 and JEF-007), whole genome sequencing and RNA-seq were performed. The whole genome of Bb ERL836 was sequenced (3,934,801,443 bp) using Pacbio sequencing and 15 contigs were analyzed. To understand the molecular mechanisms of pathogenicity in two Bb genome (before infection and after infection), next generation sequencing (NGS) was analyzed to compare transcriptomes. A result of differentially expressed gene (DEG) showed that 2,689 contigs were up-regulated and 2,619 contigs were down-regulated. Randomly selected five genes were validated to be highly up-regulated. This work can provide an understanding of the interaction between Bb and westetn flower thrips.

Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae) in forest is considered as a serious forest pest in pine trees. In this study, we collected entomopathogenic fungi from soil and assessed their virulence against the adults of the inset using a spray method in laboratory condition. Two entomopathogeinc fungi isolates showed high virulence against the adults. In the concentration of 1.0 × 107conidia/ml, the mortality rate of the adults were 53% and 60%. In semi-field condition, one isolate showed a virulence of 60% against the adult. Consequently, we confirmed the possibility of the fungal isolates in controlling the beetles. In near the future, we will investigate several factors which is possibly related to the control of the forest insect pest using entomopathogenic fungi in field conditions, given the importance of fungal formulation and practical application methods.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopenia syndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest. Therefore, this study provides a screening method to select entomopathogenic fungi, having high virulence against H. longicornis. The virulence test was conducted by dipping method with a conidial suspension (1×107 conidia/ml). As a result, cadavers of H. longicornis infected with entomopathogenic fungi were obtained by this method. Based on this results, we selected several isolates having high virulence to H. longicornis. We suggest that, selected isolates in this study can be used for the control of H. longicornis.

Entomopathogenic fungi have been known as promising candidates for biological control of insect pests. Recently, researchers consider the fungal thermotolerance in formulations and field applications. In this study, we investigated the production of thermotolerant Isaria javanica and I.fumosorosea conidia through grain-based solid cultures and exposure to light stress. As results, of the ten grain substrates, Italian millet, rice, perilla seed and sesame, rice, sorghum produced highly thermotolerant conidia in the strains. The two strains were exposed to a light stress and a heat stress. And they showed enhanced thermal stability compared to control, when exposed to 45°C for 2 hours. This work suggests that heat-resistant entomopathogenic fungal conidia can be produced by grain-based solid cultures and exposure to light stress.

Melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae) is a serious insect pest of various crops in the world. The management of this pest mainly have relied on chemical agents. However, the overuse of chemical agents for long times causes insect resistance, and negative effects on environment. Therefore, alternative control methods, such as biological control, that are less harmful to the environment and have different mode of action are needed to control this pest. In this study, we isolated entomopathogenic fungi from Korean soil, and characterized them via morphological and molecular identifications and pathogenicity check against Tenebrio molitor larvae. The isolated fungi were subjected to virulence assay against T. Palmi with a conidial suspension of 1×107 conidia/ml in laboratory conditions. This entomopathogenic fungi library would be a good resource to select effective strains for the control of T. palmi in fields.

Entomopathogenic fungi play an important natural role in regulating their insect host populations, and their ecology was also associated with plant and soil. These microorganisms have been living by reacting to insect, plant and environmental factor. The advanced bioinformatics technology such as next-generation sequencing and RNA sequencing has revolutionized in understanding of entomopathogenic fungi. Recently studies provided a lot of information on evolutionary relationships and virulence-related characteristics. We are starting to know where these microorganisms from is, and how they live in nature. The bioinformatics technology will give us further our understanding of the natural roles of these fungi in nature.

The entomopathogneic fungal colonization has become an interesting issue in ecological pest management. Plant-orientedfungi induce plant growth promotion and act as natural control agents against insect pest. Herein this work, we evaluatedthe ability of fungal colonization on bark, branches and leaves of pine tree (Pinus densiflora) to select optimal candidatesfor controlling Japanese pine sawyer (Monochamus alternatus), which is the pine wilt disease vector in Korea. As a result,some Beauveria and Metarhizium isolates could colonize on the substrates of P. densiflora and showed pathogenicity againstM. alternatus. This result suggests that the selected entomopathogenic fungi are possibly candidates for long-term managementof M. alternatus in pine tree forest.

Japanese pine sawyer, Monochamus alternatus HOPE (Coleoptera: Cerambycidae) are considered as important pine forestinsect pests in several countries which transmits the pine wood nematode, Bursaphelenchus xylophylus. In this study, wetried to control Japanese pine sawyer using entomopathogenic fungi to prevent the spread of pine wilt disease in Korea.Several entomopathogenic fungi were exposed to the adults of Japanese pine sawyer and mycosis was observed in themembranous cuticles after 6 days. In particular, two isolates showed high virulence against the adults. The selected 2isolates were sprayed to the adults at 1×105, 1×106 and 1×107 conidia/ml. In the concentration of 1×107 conidia/ml, thesurvival rates of insects were 40% and 47%, respectively. Based on these results, the possibility of controlling the Japanesepine sawyer was confirmed.

Chemical pesticides have been used for persimmon pest control, however the overuse caused insect resistance andenvironmental toxicity. Herein this work, we investigated the biological potential of eco-friendly organic pesticides (biologicals)and entomopathogenic fungi in controlling persimmon pests, Stathmopoda masinisaa and Riptortus pedetris. A botanicalmixture of Chinese scholar tree, goosefoot and subtripinnata extract, Bacillus thuringesis NT0423 and one isolate of Beauveriabassiana showed high insecticidal activity against the pests in laboratory conditions. The selected botanical extract andmicroorganism were combined with a chemical pesticide (buprofezin+dinotefuran WP) and were assessed their controlefficacy in persimmon fields. Compared to the three times of chemical spray, the combination of more than one timeof chemical spray and the selected biologicals controlled the persimmon pests with an acceptable levels. Consequentlyone time of biological spray can be integrated to the spray program where only chemicals are strongly considered.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopeniasyndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest.We evaluated three bioassay methods, such as spray, dipping and dropping. As a result, the dipping method was themost efficient way to measure the fungal virulence. Finally, we constructed a Haemaphysalis longicornis pathogenic fungallibrary to further facilitate the resources to be used as potential biological control agents. A conidial suspension of 1×107conidia/ml was exposed to the ticks by dipping. We confirmed that some isolates were highly virulent to the ticks. Thisresults indicated that some fungi could be used to effectively control the Haemaphysalis longicornis.

Many entomopathogenic fungal isolates received interests for controlling insect pests. But very few numbers of isolatesreached agricultural fields as commercial products. In this study, we investigated the feasibility of grain substrates forconidial production, focusing on conidial productivity and thermotolerance. Next to efficiently use the produced conidiafor spray application, conidia need to be separated from the grain substrates. Isaria javanica and I. fumosorosea conidiawere solid-cultured on ten kinds of cereal grains. Solid-cultured grains were packed in a screen bag and immersed inwater with a variety of surfactants at 100 ppm, such as Silwet, CO-2.5, CO-12, LE-7, PE-61, TDE-3, Tween 20 or Tween80. High levels of conidial release were observed on LE-7, TDE-3 and Tween80 treatments. This work suggests thatthe screen bag can be used to separated conidia from the cultured grains with the help of surfactants.

A genus of entomopathogenic Beaveria bassiana has been widely used in pest management, however little studies havebeen given to its virulence-related genes. To identify the roles of virulence genes, AtMT-induced random mutants weregenerated and followed by localization study with TAIL-PCR. Two genes, Complex I intermediated-associated protein30 (CIP30) and Autophagy protein 22 (Atg22), were predicted as virulence-related genes in B.bassiana JEF-007. To validatethe a possible relationship between two genes and fungal virulence, hpRNAi was performed. A hpRNAi plasmid wasconstructed as a model system to knock down of egfp gene in egfp-expressing B. bassiana transformant. A Real-TimePCR showed the knock down of egfp gene expression via hpRNAi. The CIP30 and Atg22 genes in B. bassiana willbe validated by the established RNAi technique. This work can be a strong platform for the functional genetics in B.bassiana.

Entomopathogenic fungi, widely distributed in nature, have been developed as biopesticides of agricultural pests. However, little is known about isolation method for obtaining a variety of highly virulent strains from the soil. In this work, the previously established method using mealworm (larvae of Tenebrio molitor) as a baiting system was used to efficiently isolate fungal strains from soil, and the isolated fungal library in Korea was established. To isolate entomopathogenic fungi, soil samples from 10 regions of Korea such as Iksan, Imsil, Gunsan, Yeosu, Suncheon, Gwangyang, Jinju, Hamyang, Boseong and Jangseong were collected. Of the collected soils, 21 and 34 isolates of entomopathogenic fungi were isolated from Iksan and Gunsan, respectively. The isolates were classified as genera of Metarhizium, Beauveria and Isaria. This work can suggest an effective insect pathogenic fungal isolation method

Entomopathogenic fungi have been known as promising candidates in biological control. However, a large number of conidia with higher stability should be produced for industrialization. In this work investigated the possibility of several grains for the production of thermotolerant Isaria javanica and I. fumosoroseus conidia. The fungal conidia were produced on ten grains for 14 days in Petri dish conditions. Of the ten granular substrates, millet, perilla seed and barley showed the high conidial production, 15.7 ± 8.4, 17.3 ± 5.2 and 13.5 ± 1.9 × 109 conida/g in I. javanica and 15.9 ± 0.6, 15.0 ± 0.5 and 15.0 ± 0.7 ×109 conida/g in I. fumosoroseus, respectively. However, the two strains showed relatively low thermal stability regardless of substrates, when exposed to 45℃ for 1, 2 and 3 hours. This work suggests a possible substrate for conidial production of entomopathogenic fungi with low cost.

Entomopathogenic fungi are expected to play a role as great biological control agents in the global bio-pesticide market in the future. The first step is the collection of fungal isolates and it should be a platform for the development of highly effective biopesticides. In this work, we constructed a fungal library using a mealworm pathogenecity-based fungal collection method and further characterized some isolates with high virulence. A phylogenetic tree was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, virulence against several insect, stability of conidia for heat, production of biologically active materials, such as enzymes. This work reports an attractive entomopathogenic fungal library including the information of effective isolates in pest management.

Many turfgrass pests, Ectinohoplia rufipes, Exomala orientalis and Popillia quadriguttata, cause serious damage to grass fields in Korea. In this study, control of turfgrass pests using entomopathogenic fungi were investigated using Protaetia brevitarsis seulensis as an alternative insect. In a preliminary experiment, some Metathizium sp. showed ca. 60% virulence after 7 days exposure to the entomopathogenic fungi. Compared to the contact-exposure, overall a spray treatment showed lower control efficacy against P. brevitarsis seulensis. Furthermore, mortality of various developmental stages of P. brevitarsis seulensis was investigated under laboratory conditions. This work suggests that when treated with fungi mortality of P. brevitarsis seulensis was closely related with developmental stage. These results provide a basic information for the control of P. brevitarsis seulensis using entomopathogenic fungi

Entomopathogenic fungi are potential pesticides as biological control agents. However, many previous studies have focused on basic researches to control pest using insect-pathogenic fungi. Herein, the molecular characteristics of insect defense mechanisms against fungal pathogens and fungal pathogenicity to insects were investigated. The whole genome of Beauveria bassiana (Bb) JEF-007 was sequenced (36,538,394 bp) using Pacbio sequencing. Based on this data, the transcriptomes of bean bug and fungus were analyzed using RNA-seq. In the transcriptome analyses, six and five genes were validated to be involved in the fungal pathogenicity and insect defense, respectively. This work can provide an understanding of the insect-pathogen interactions

본 연구는 퇴비에 함유된 영양소 및 중금속 함량을 파악하고 상추 재배시 퇴비의 적정 시용 비율을 알아보고자 수행되었다. 실험을 위해 두가지 퇴비를 이용하였다. 첫 번째 퇴비는 미완숙 퇴비(CA)이며 두 번째 퇴비는 시중에서 판매되고 있는 완숙 퇴비(CB)이다. 각각의 퇴비는 인공토양을 0%, 25%, 50%, 75%로 혼합하여 사용하였다. 50%와 75%의 비율로 혼합한 CA의 pH는 각각 5.39, 5.50으로 측정되었으며 약 산성으로 나타났다. CA 및 CB를 75% 비율로 혼합할 경우, 총 탄소 함량은 각각 14.5%와 6.5%로 다른 비율의 퇴비에 비해 높았고 대조구에 비해 총 질소와 인 농도가 유의하게 증가하였다. 총 탄소함량은 CA퇴비를 인공토양에 75% 혼합한 실험구에서 가장 높게 나타났다. CA는 CB와 비교하여 퇴비화율, 질소, 인의 농도가 크게 증가하였다. CB 75% 혼합한 실험구에서 구리(128 mg kg-1), 아연(260 mg kg-1), 납(0.32 mg kg-1), 카드뮴(0.48 mg kg-1) 의 함량은 다른 혼합구에 비해 가장 많은 증가하였다. 특히 비소는 CA퇴비를 25% 혼합한 실험구와 CB퇴비를 75%, 50% 혼합한 실험구에서 가장 높았다(6.69 and 6.28 mg kg-1). CA실험구 중에서 상대적으로 낮은 염분 및 중금속 함량을 함유한 CA 25% 혼합한 실험구는 상추의 성장속도 및 엽면적 등이 CB에 비해 낮게 측정되어 최적의 성장조건은 아닌 것으로 사료된다. 따라서, CA를 사용하여 상추재배에 이용할 경우, 더 낮은 농도의 CA를 이용하는 것이 적당할 것으로 판단된다.