It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.

Host defense against pathogen invasion highly relies on immune defense machinery that is controlled by the nuclear factor-κB (NF-κB) of transcription factors. The Toll pathway are well known as an insect innate immune mechanism to protect host itself from invaded pathogens. Basically, in the edible insect, Tenebrio molitor, the Toll pathway is primarily activated by polymeric Lys-type peptidoglycans (PGNs), and components of fungal cell walls, β-1,3-glucan. Based on the current studies, the tremendous study has been focused on recognition and subsequent activation of spätzle in haemolymph, hence, there is a grave gap for intracellular event. Herein, in order to understand intracellular event of Toll signaling pathway, the Dorsal gene were identified. Moreover, domain analyses of TmDorsal2 gene indicate that there are two major domains such as Rel homology domain (RHD), ig-like, plexins, and transcription factors (IPT) domains. Based on the achieved results, TmDorsal2 mRNA was highly expressed in 1-day old pupa. Furthermore, TmDorsal2 was highly expressed in Malpighian tubules and fat body in last instar larvae (LL), and likewise mainly expressed in Malpighian tubules during adult 5-day old period, also the lowest expression of TmDorsal2 was observed in gonads. Moreover, TmDorsal2 mRNA levels after infection with E. coli appreciably went up at 6 and 9h time points. To investigate the effects of TmDorsal2 RNAi on larval susceptibility against various pathogens namely E. coli, S. aureus or C.albicans, dsRNA of TmDorsal2 has been synthesized the larvae dissected after 24h. As a result, TmAttacin1a, 1b and 2, TmDefencine1 and 2, TmTenecin1, 2, 3 and 4, TmCecropin2, TmColeoptericin1 and 2, Thaumatin-like protein 1 and 2 markedly reduced in the gut after injecting all mentioned microbes. In contrast, TmTenecin 2, Thaumatin-like protein 1 and 2 strikingly increased after microbe injection in the fat body. Interestingly, the most AMPs gene expression in whole body experimental case were upregulated. On the horizon, we will investigate effects of TmDorsal1 RNAi on larval susceptibility against various pathogens. Taken together, our studies may aid to understand insect innate immunity.

Gonggeom-ji is designated wetland protected area by the Ministry of Environment of Korea in 2011 because of it’s high biodiversity and historic value. It contains a reservoir, paddy and forest site which has diverse niches for insects. Three transect lines of 50m were designated. Data were collected in May, October, November in 2017 represented as seasons for spring, summer and autumn respectively. Quantitative methods were conducted along each transect line by sweeping and pitfall trap. Based on this study total of 1079 individuals of insect fauna were collected, representing 170 species in 60 families and 8 orders. The highest diversity, richness and evenness index were observed in the forest site in May(4.77, 8.6 and 0.91 respectively) and the highest dominance index was observed in the forest site in November(0.64). The highest similarity index was observed in the reservoir site in May and August(0.519).

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

The objectives of this study was to evaluate the degradability and digestibility of crude protein (CP), rumen undegradable protein (RUP), and individual amino acids (AA) on six by-product feedstuffs (BPF) (rice bran, RB; wheat bran, WB; corn gluten feed, CGF; tofu residue, TR; spent mushroom substrate from Pleurotus ostreatus, SMSP; brewers grain, BG) as ruminants feed. Three Hanwoo steers (40 months old, 520 ± 20.20 kg of body weight) fitted with a permanent rumen cannula and T-shaped duodenal cannula were used to examine of the BPF using in situ nylon bag and mobile bag technique. The bran CGF (19.2%) and food-processing residue BG (19.7%) had the highest CP contents than other feeds. The RUP value of bran RB (39.7%) and food-processing residues SMSP (81.1%) were higher than other feeds. The intestinal digestion of CP was higher in bran RB (44.2%) and food-processing residues BG (40.5%) than other feeds. In addition, intestinal digestion of Met was higher in bran RB (55.7%) and food-processing residues BG (44.0%) than other feeds. Overall, these results suggest that RB and BG might be useful as main raw ingredients in feed for ruminants. Our results can be used as baseline data for ruminant ration formulation.

In recent years, pavement distresses have been caused by diverse factors such as spalling, deterioration of repaired sections, blow-up, and alkali aggregate reaction due to changing climate environment of a concrete pavement and its construction and maintenance conditions (supply of materials, increase in use of de-icers, etc,). As a leading repair method for deteriorated concrete pavements, partial-depth repair is implemented in accordance with guidelines of material properties for joints of a concrete pavement and field application evaluation systems, but still some of the repaired sections become deteriorated again at an early stage due to poor construction quality and failure of response to environmental impacts. Distresses that can be corrected with partial-depth repairs are largely divided into those of repair materials and of the existing pavement bonded to repair materials, and combined distress of repair materials and the existing pavement. Although re-repair methods should be different by distress type and scale than conventional pavement repair methods, appropriate repair methods and guidance for re-repairs have not been in place so far, and therefore currently, re-repair practices follow the existing manual of partial depth repairs. Therefore, this study evaluated concrete bond characteristics by removing method and repair scope for an experimental section of frequently distressed pavements to determine a re-repair scope and method for deteriorated partial depth repair sections of concrete pavement, the number of which has increased over time.

This research was relative humidity (65, 75, 85 and 95%) growth characteristics of the oak mushroom (Lentinula edodes) 'Nongjingo’. At the relative humidity was 65% in growth chamber. On this condition the moisture content of medium surface was 60.5%. That was lower than other treatments. That condition makes Pileus was 77.1% and stipe was 65%. Among the growth characteristics according to the relative humidity, the pileus diameter at 75% relative humidity were 45.5mm that largest than other treatment and pileus thickness at 75% relative humidity were largest than other treatment. The stipe thickness at 65% relative humidity were 14.8mm that lagest than other treatment. The number of available stipes at 95% relative humidity were 13.3peace/2kg that largest than other treatment. The comparison yield ability of 1 cycle was 104.2g that result in largest than other treatment at 95% relative humidity. This result was associated to the relative humidity of fruiting bodies, the lower relative humidity result in the lower of yield. However, the lower relative humidity result in the higher number of whago, Especially, at the treatment of 65% relative humidity, all the fruit bodies were produced as Whago (dry). Therefore, the higher relative humidity influenced in the higher yield but produced water mushroom. That result was thought that ‘Nongjingo’ would be possible to produce high quality mushroom by adjusting the relative humidity appropriately.

The green peach aphid, Myzus persicae, is one of the most important pests of vegetable crops worldwide. This species cause direct damage by feeding on plant nutrients and indirect damage as transmits many virus vectors. The control of aphid has relied on the use of chemical insecticides and their intensive use over many years has led to the development of resistance. To surmount or avoid these problems, we screened the entomopathogenic fungi against green peach aphid and evaluated their virulences. Several entomopathogenic fungal isolates were selected with high virulence to green peach aphid. Additionally, the antimicrobial activity of the selected fungal isolates was tested and analyzed to phytopathogens. Consequently, these entomopathogenic fungi would be used effectively for dual control agents for the green peach aphid and phytopathogen.

Earthworms, especially Eisenia andrei, are cultivated for a variety of purposes including waste disposal and compost production. In this study, bacteria from earthworm gut were cultured and the dominant species identified. Subsequently, we isolated bacteriophages able to lyse the isolated gut bacteria. Two dominant genera of gut bacteria, Aeromonas sp. and Citrobacter sp. were identified by using MALDI-TOF MS analysis and a library was constructed to find lytic phages. Phage EF1 showed lytic activity for C. freundii and two Citrobacter isolates, C. braakii and C. murliniae. These 3 species have similar sensitivities to EF1. Several aspects of the life cycle of EF1 were investigated by using C. freundii under optimal growth conditions. EF1 infects C. freundii with a moderate latent period, approximately 25 min, and a large burst size averaging 5 × 109 per infected cell. Moreover, the antimicrobial activity of EF1 was well maintained under diverse conditions including a broad temperature range of 40°C to 50°C and a wide pH range of 4 to 11. In conclusion, the results indicate that earthworm casting contain a wide range of bacteria species, for which there are various corresponding bacteriophages.

We developed a simple molecular detection tool that rapidly and accurately identifies Spotted wing Drosophila (SWD) without sophisticated instruments or expertise. We first identified a gene that is present in the SWD genome but not in that of any other insect species. Then, we developed the loop-mediated isothermal amplification (LAMP) assay, which was designed based on genomic DNA of SWD specific gene. This LAMP assay detected only genomic DNA from SWD—not from Drosophila melanogaster. In addition, this assay could detect genomic DNA in SWD geographical strains collected from 8 different locations in Asia, Europe, Hawaii, and the USA. Our LAMP assay could be a useful detection tool for identifying SWD rapidly in the field. This work was carried out with the support of the Cooperative Research Program for Agriculture Science & Technology Development (Project No: PJ0116302016).

To obtain information on the sanitary of indoor environment in greenhouses used for shiitake cultivation, bacteria associated with larvae and imagoes of Sciaridae flies, pest to shiitake mushroom, were isolated and identified. A total of 1,048 bacterial colonies were isolated from the flies’ larvae and 984 bacterial colonies were isolated from the flies’ imagoes. Based on molecular analysis of the 16S rDNA sequence, Achromobacter xylosoxidans and Tetrathiobacter kashmirensis of β-proteobacteria, Enterobacter asburiae and Raoultella ornithinolytica of γ -proteobacteria, Curtobacterium sp. and Microbacterium thalassium of Actinobacteria, and Penibacillus taichungensis of Firmicutes were identified from the colonies of the flies’ larvae. While, Bacillus megaterium, B. thuringiensis, Lysinbacillus sphaericus and L. fusifomis of Firmicutes, Microbacterium thalassium and Citricoccus parietis of Actinobacteria, and Enterobacter asburiae of γ-proteobacteria were identified from the flies’ imagoes. Some of the isolated bacterial species were known be human pathogens. Overall, the results of this study suggested that mushroom fly carrying human pathogenic bacteria is one of sources impact on the sanitary of indoor environment of greenhouses used for shiitake cultivation.

Even though plant protections using chemical pesticides have several advantages, non-specific toxicities to other beneficial insects and humans and rapid development of tolerance and/or resistance of target pests to chemicals are major disadvantages. Recent researches suggested that using double stranded RNA (dsRNA) could be species specific and environmental friendly pest management protocols. However, efficiency of dsRNA treatments are known to be variable according to its application methods. For example, injections of dsRNAs to pests were known to be effective in all species. However, efficiencies of oral application of dsRNAs were known to be dependent on species. Thus, development of tools that could enhance the efficacy of orally treated dsRNAs are utmost important for widening usages of dsRNAs in plant protection. Recently, we found that the efficacy of oral treated dsRNAs to target pests could be enhanced by nano-technologies. I will show how applying nano-technology to dsRNAs enhance the efficiency of dsRNAs. (This work was carried out with the support of the Cooperative Research Program for Agriculture Science & Technology Development (Project title: Studies for biological characteristics of and control methods against the migratory locust, Locusta migratoria, Project No: PJ011630042016), Rural Development Administration).

Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in pigs with high mortality. PEDV is belong to Coronavirus, enveloped, single-stranded, positive-sense RNA virus. PEDV particles were composed of four structure proteins such as a glycosylated peplomer (spike, S) protein, envelope (E), glycosylated membrane (M) protein, and unglycosylated RNA-binding nucleocapsid (N) protein. Many of previous studies talk about this four structure proteins have a great potential to diagnosis and prevent PEDV. In this study we investigated expression of these structure proteins using the bacterial and baculovirus expression system. In bacterial expression system, our results showed that structure proteins fused polyhedrin and intein gene were expressed higher than non-fusion structure proteins. The expressed fusion proteins were used to immune mice for generating a polyclonal antibodies. In baculovirus expression system, co-infection of insect cells with these four recombinant baculoviruses led to self-assembly of virus-like particles as demonstrated by Transmission electron microscopy. They were confirmed by western blot analysis using pre-made polyclonal antibodies. Finding in this study may provide important information for vaccine and diagnostic development.