As ubiquitous organisms entomopathogenic fungi may natually occur within environment including phylloplanes and soil or deposite and survive for a while after spraying a fungal suspension to control insect pest. The existence of pathogenic fungi is a threat to arthropods foraging for food and oviposition sites. The detection and avoidance of pathogens is important for host survival, longevity and ultimate fitness. Although entomopathogenic fungi are ubiquitous worldwide and act as common natural enemies of many invertebrate species, very few studies have conducted abiut the detection and avoidance of pathogenic fungi by insect pest. We have studied that an insect pest can recognize a high virulence fungi on crop surface, avoid oviposition and don’t enter the field treated their pathogen.

Entomopathogenic fungus is a useful control agent to sucking type insect such as whitefly and aphid. The fungi are influenced by some environmental factors such as relative humidity, temperature and UV and cause slow and fluctuation in pest control efficacy. Especially, UV kills conidia or spores of entomopathogenic fungi and a mycopesticide using fungi has short control period in field. UV intensity changes from season to season. Survival rate of entomopathognic fungi treated may differ from seasons and will show different control efficacy. Therefore, we conducted a study to estimate the persistence of an Isaria javanica isolate, which was already reported as sweet potato whitefly control agent, in potted greenhouse soil planted different crops. The number of survival spore decreased gradually and differ from seasons.

Entomopathogenic fungi are natural enemies of insect pests and contribute to the natural regulation of their host populations. These fungal group are often used as active ingredients for microbial insect pest control. In addition, the potential antimicrobial effect by entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have recently been reported against fungal plant pathogens. Dual microbial control effects with entomopathogenic fungi against both aphids and cucumber powdery mildew had reported in Canada. In our previous studies we conducted bioassay with entomopathogenic fungi to develop dual microbial control agent which can control both aphid and fungal plant disease. We selected an Beauveria bassiana isolate which has high dual control effects against both cotton aphid, Aphis gossypii and sclerotinia rot, Sclerotinia sclerotiorum. In this study, we have tested the dual control efficacy of the B. bassiana isolate against cotton aphid and sclerotinia rot on whole potted cucumber plants. We found that the B. bassiana isolate protected the plant from cotton aphid and sclerotinia rot under laboratory condition.

Insect pests have been a serious problem over many years and remain a major threat for food production. Although chemical pesticides are major pest control strategies, use of microorganisms such as entomopathogenic bacteria, fungi, viruses and nematodes have continuously increase last few decades to minimize the use of agrichemicals. According to BBC research, the global biocontrol market was about $2.1 billion in 2011, and this is expected to rise $3 ~ 4 billion by 2017. Over 50 entomopathogens are commercially produced and used augmentatively as microbial pesticides. About 175 biopesticide active ingredients and 700 products have been registered worldwide. Bacillus thuringiensis (Bt), Beauveria bassiana, Metarhizium spp., nuclear polyhedrosis virus and Steinernema spp. are the most popular control agents used in plant protection. Among the microbial control agents Bt products have more than 50% of market share. In Korea, only 13 environmentally-friendly crop protectants were registered for insect pest control in 2015. Market share is very low and has grown slowly. We will discuss how we can expand the market with our techniques.

경기도 8개 지역에서 2010년부터 2012년 동안 식균성인 노랑무당벌레의 발생기주를 조사한 결과, 흰가루병에 감염된 12종의 식물에서 관찰 이 되었다. 특히 가장 밀도가 높았던 배과원에서 노랑무당벌레는 7월 상순부터 11월 상순까지 발견되었다. 식균성인 노랑무당벌레의 장내에서는 흰가루병 균사나 포자 외에 다른 먹이의 흔적이 발견되지 않았고, 알과 번데기를 제외한 전 발육단계에서 균을 섭식하는 특성을 볼 때 절대적 식균성 곤충으로 생각된다. 25℃에서 오이 흰가루병균을 섭식한 노랑무당벌레의 발육기간은 알, 유충, 번데기, 성충이 각각 3.9, 10.4, 4.1, 37.7일 이었고, 발육단계별 오이 흰가루병 섭식량은 45.6, 144.4, 372.2, 628.1, 473.7 mm 2로 4령, 성충, 3령, 2령, 1령 순으로 많았다. 본 연구를 통해 노랑무당벌 레의 오이 흰가루병에 대한 섭식능력을 바탕으로 향후 유용 토착천적으로써 대량사육기술, 저독성 약제 선발 등 작물 흰가루병 종합방제기술(IPM) 에 대한 연구가 필요하리라 사료된다.

This study was conducted to evaluate the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To determine the movement of degradable HGC nanoparticles in the body, 20 mg/kg of lutetium177-labeled HGC (Lu177-HGC) with the size ranging from 320 to 400 nm was injected intravenously into ICR mice, and the amount of radioactivity remaining in blood and several organs was measured at various time points during the period of 5 days. In the pharmacokinetics analysis using the Lu177 radioisotope, the free Lu177 was mainly distributed and accumulated in the order of kidney>liver>lung at 1 day after the injection of the radioisotope. However, the Lu177-HGC showed a high distribution of nanoparticles in the order of liver>spleen>kidney during the experimental period of 5 days. These results would provide a basic pharmacokinetics for the use of HGC as a drug carrier in drug delivery system.

The ultimate goal of this study is to assess the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To understand the movement of degradable nanoparticle HGC in the body, we intravenously injected a dose of 20 mg/kg of Cy5.5-labeled HGC with size ranging from 320 to 400 nm into ICR mice, and measured the amount of fluorescence remaining in blood and several organs at various time intervals. In blood, the level of Cy5.5-labeled HGC was the highest at 15 min, then after 30 min it decreased rapidly and reached a plateau form 30 min to 28 days. In the tissue we confirmed the presence of nanoparticles at high levels in the order of kidney>liver>submandibular gland until 28 days after injection. However, we did not find the presence of the particles in the brain or testes. These results will provide basic information on HGC as a drug delivery agent.

Skin is the outermost organ and acts as a barrier between the organism and environment. Skin protects the organism from environmental insults, such as chemicals, pathogens, and UV light. Much of the protective function of skin is dependent on the epidermis, a multi-layered epithelium that is composed of various cell types such as keratinocytes and melanocytes. Keratinocytes produce protective components through a sophisticated differentiation process. Disturbance of keratinocyte differentiation is related to several skin diseases such as psoriasis and atopic dermatitis. In this study, we prepared extract of combined medicinal plants (ECMP) consisting of Taraxacum platycarpum H. Dahlstedt, Heartleaf Houttuynia, Glycyrrhiza uralensis Fischer, and root bark of Ulmus davidiana. We demonstrated that ECMP enhanced keratinocyte differentiation and barrier functionality using an in vitro cell culture system and in vivo animal test. Treatment of cultured keratinocytes with ECMP resulted in induction of keratinocyte differentiation, as evidenced by increased differentiation markers such as involucrin, loricrin, and filaggrin. In line with these results, ECMP decreased proliferation of keratinocytes cultured in vitro. ECMP applied topically to tape-stripped mouse skins accelerated reduction of transepidermal water loss (TEWL), indicating fast recovery of barrier function. Immunohistochemistry showed that ECMP increased the filaggrin level in tape-stripped mouse skins. These results suggest that ECMP may be applicable for keratinocyte differentiation-related skin diseases.

Biological properties of antimicrobial peptides (AMPs) of hemimetabolous insect are poorly characterized in innate immunity field. To investigate the biochemical properties of hemimetabolous insect’s AMPs, we purified the pyrrhocoricin-like AMP from the hemolymph of Riptortus pedestris and then named as riptocin. We successfully determined the primary protein structure and its cDNA sequence. Interestingly, the determined cDNA revealed that riptocin precursor is composed of 12 repeating units of active riptocins, which implied that riptocin precursor might require to be processed to generate active riptocins by several unidentified processing enzymes. In order to characterize the bio-processing mechanisms of riptocin precursor, we generated the antibody against active riptocin. Using quantitative PCR and Western blot analyses, we showed that gene of riptocin was started to express from the fatbody after three hours post bacterial infection. To address our hypothesis that active riptocin is generated from riptocin precursor by several processing enzymes, we need to obtain the riptocin precursor. Currently, we are expressing the recombinant riptocin precursor using in vitro translation system. Meanwhile, we investigated whether naive hemolymph (naive HL), which may contain precursor riptocin, can generate active riptocin when riptocin precursor was co-incubation with bacteria-challenged hemolymph (active HL), which may contain all processing enzymes. Actually, when naive HL was incubated with active HL, antimicrobial activity was dramatically increased, suggesting that processing enzymes in active HL may induce processing of riptocin precursor to generate active riptocins.

A choice in temporal discounting is often made on one preferred option between a sooner but smaller reward and a later but larger reward. In temporal discounting, a consumer undergoes an evaluation process concerning the trade-offs between time delay (i.e., a choice between sooner and later receipts) and the amount of reward (i.e., a choice between smaller and larger rewards). It has been evidenced that a surprisingly large number of consumers is inclined to choose a sooner option with a lower economic value. Those consumers tend to apply an excessive temporal discounting rate to the time delay and as a result, the sooner option with lower economic value stands out to be more attractive. However, there is another group of consumers who deliberately promote the long-term perspective in the evaluation process. Those consumers show a high tendency of choosing a later option with greater economic value. A key factor affecting the evaluation process is the sensitivity to time delay a consumer entails. The sensitivity to time delay could systematically affect the representations of hedonic pleasure being projected at different time delays and is the basis of individual differences in hedonic orientation. An anhedonia, a person with the ability to control physical pleasure, is likely to be less sensitive to time delay and apply a lower discounting rate for delayed rewards because he processes the presently available pleasure less intensively than do ordinary consumers. Thus, anhedonic consumers are more likely to choose a delay reward in temporal discounting choices. In a famous fable by Aesop, “The Ant and the Grasshopper” suggest a possible linkage between hedonic orientation and temporal discounting tendency. The story goes that the Grasshopper enjoys whatever is on its hands and does not give much thought to what is going to happen in the future, while the Ant takes on the hardship denying the current pleasure as he plans for the difficult winter time. We conducted an experiment to investigate the relationships among individual differences in hedonic orientation, and the behavioral choices in temporal discounting trials by examining the EEG theta (4-8 Hz) correlates. A hedonic orientation questionnaire was developed using twenty selected items from Physical Anhedonia Scale, Barratt Impulsiveness Scale and Impulsive Buying Scale. Using the questionnaire, the hedonic orientation of fifteen participants was analyzed. Among them, five participants were classified into anhedonia (Ant-like, AN) group and ten participants were into hedonic pleasure dependent (Grasshopper-like, GH) group. Participants conducted ten temporal discounting trials and, in each trial, a choice was made between two options, either a smaller reward today (the “now” option) or a greater reward to be received after a specified time delay (the “future-oriented” option). The absolute theta activations while viewing a future-oriented option in each trial were analyzed as the neural correlates of temporal discounting. The results indicated that the hedonic orientation of participants had a high prediction power of not only behavioral choices in temporal discounting but also EEG theta activations in the frontal brain region. In analyzing temporal discounting tendencies, the discounting rate of AN group was found to be lower than that of GH group. In addition, EEG theta analysis shows two interesting activation patterns in the frontal region. First, AN group showed higher frontal theta activations than GH group indicating its deliberate engagement to future-oriented options. Second, theta activations in the Dorsolateral Prefrontal Cortex (DLPFC), more specifically AF3 and AFz, were negatively correlated with the discounting rate of participants. Neuroscience researches often refer that DLPFC is associated with deliberate and analytic information processing. Therefore, the high theta activations in DLPFC could be considered as a distinctive neural indicator of high anhedonia traits and low temporal impulsivity.

The aim of the present research is to explore the neural features of intertemporal choice. Intertemporal choice concerns the phenomenon of temporal discounting which demonstrates consumers’ tendency to devalue a future reward as the receipt of money reward is delayed further distantly into the future. It is also called the ‘present bias,’ a special kind of attachment consumers place to immediate a cash reward which consumers are willing to take even at a discount, instead of saving it for the future. The urge to choose an immediate reward at some discounting rates, i.e., temporal impulsivity, is deeply embedded in our instinct. On the other hand, the choice for a delayed option is made as a result of effortful and deliberate cognitive control, by suppressing the urge for instant gratification and shifting focus to the long-term gain. Neuroscience researchers have found the prefrontal cortex as a key brain region behind deliberate and analytic decision-making such as reward delay. The experimental paradigm used in this study adopted from Kirby, Petry, & Bickel (1999). There were ten intertemporal investment choices representing six different levels of discounting rates. Participants (N=21) were given the two investment options, as “take it now” and a “save for later” options, and they were asked to indicate a preferred option. The brain waves were recorded using a 32 channel EEG system. We conducted noise control by using Independent Component Analysis (ICA) in EEGLAB and analyzed the Event-Related Potentials (ERP) particularly focusing on the phase when each participant was examining the future-oriented “save for later” option. The behavioral responses were divided into two groups, one in which the present option was selected and the other in which a future-oriented “save it for later” option was selected. Custom-written Matlab codes were used to compare the differences in ERPs between the two choice responses. A greater positive component of P3 after 300 milliseconds at the onset of the future option was observed in the fronto-parietal regions when the “save it for later” options were chosen compared to when the “take it now” choices were made. Hence, during intertemporal choice, the fronto-parietal regions are likely to play a key role in enabling the long-term perspective and consequently, precipitate the choice of a future-oriented deal. The P3 signal across the fronto-parietal regions may potentially be a proximal indicator of a future-oriented choice during intertemporal financial decision-making.

Beet armyworm, Spodoptera exigua is difficult to control using chemical insecticides because of the development of insecticide resistance. Various control agents may use for eco-friendly beet armyworm managements, Entomopathogenic fungus is one of the promising control agents as an alternative to chemical control agent. At previous study we isolated a number of entomopathogenic fungi from soil samples and selected three entomopathogenic fungi, Metarhizium anisopliae FT83, Paecilomyces fumosoroseus FG340 and Metarhizium anisopliae FG344 which had high virulence against larva of beet armyworm. To select an isolate having high virulence at high temperature against S. exigua, thermophilic pest, we conducted bioassay at five different temperatures(15, 20, 25, 30, 35℃). These three isolates showed the high mortality and low LT50 at 30℃. To select substrate for mass production these isolates were inoculated(5×10⁴/㎖, 20㎖) at barley, unpolished rice, wheat bran and rice bran and incubated for 14 days at 25℃. Wheat bran showed highest conidia production of M. anisopliae FT83 and P. funosoroseus FG340 and barley was suitable for M. anisopliae FG344.

Various insect pests infest crops including vegetables and ornamentals during crop production. Chemical pesticides have generally used until recently to control pests. Many pests are very difficult to control using organic pesticides because of the development of insecticide resistance and their rapid population increase. Entomopathogens are the promising alternative control methods. Entomopathogenic fungi are good candidates to control sucking insects such as aphid, whitefly and mite because the fungi can infect without ingestion. Conidia or blastospores of fungal entomopathogens are sprayed onto target plant and/or insect, adhered on insect surfaces, and penetrated into hemocoel through insect cuticle. Then the fungus utilize insect nutrients and kill the host insect. During this process, fungus was influenced by environmental conditions such as temperature, relative humidity and UV light. These are causing slow mortality and preventing wider application and use of mycopescitide using entomopathogenic fungi. In addition, control efficacy with fungal entomopathogen differed fungal isolate and host insect. Therefore, we need to study selection of high virulence isolate, mass production, formulation and application techniques to develop mycopesticide.

Emergence of resistant two-spotted spider mite (TSSM) can induce the over usage of standard amount of acaricides and result in various side effects. Rapid resistance monitoring is essential step for the efficient management of resistant populations by enabling the selection of appropriate acaricides. Here, we evaluated the 19 acaricides to determine its suitability for residual contact vial bioassay (RCV) by using PyriF strain as a reference. Twelve acaricides (Amitraz Abamectin, Bifenthrin, Bifenazate, Chlorfenapyr, Cyenopyrafen, Cyflumetofen, Endosulfan, Fenothiocarb, Monocrotophos, Omethoate and Tebufenpyrad) revealed the dose-dependent mortality within 8 h, whereas other remaining acaricides (Dicofol, Etoxazole, Fenbutatin oxide, Fenpyroxymate, Flufenoxuron, Spiromesifen and Pyridaben) did not. This finding suggests that the application of RCV method is limited depending on the mode of action and physicochemical properties of each acaricide. Resistance levels to 12 acaricides were determined for four field populations of TSSM by using RCV diagnostic kit. All TSSM populations showed the highest sensitivity to cyflumetofen, indicating that it would be most effective in controling field populations. RCV diagnostic kit would enable to provide crucial information for choosing the most appropriate acaricides in the field.

Beet armyworm, Spodoptera exigua are difficult to control using chemical insecticides because of the development of insecticide resistance. For eco-friendly beet armyworm managements, various control agents are required. Entomopathogenic fungus is one of promise control agents as an alternative to chemical control agent. We isolated entomopathogenic fungi from soil samples of suwon by insect-bait method using Galleria mellonella and conducted bioassay to larva of beet armyworm. As a result of bioassay isolate FG274, FG340, FG344 had high virulence as 100% against second instar larva of S. exigua. To identify the fungus isolates, their’s morphological characteristic was observed and ITS of 18srRNA was sequenced. ITS sequence of FT274, 340, 344 were highly matched (100%) to that of Beauveria bassiana, Paecilomyces fumosoroseus, Metarhizium anisopliae. To investigate the optimal concentration, three isolates were sprayed at three different concentration(1×106 ,107 and 108 conidia/㎖) in laboratory conditions. 나타내었다.

Use of conidia or spores of entomopathogenic fungi are influenced by environmental conditions such as temperature and relative humidity and caused slow and fluctuation of mortality. In addition, although the fungi have the advantage of a restricted host range, this specificity is one of the limiting factors for their use. These factors are preventing wider application and use of these biocontrol agents. To mitigate such problems we selected an Beauveria bassiana Bb08 which kill green peach aphid with its liquid culture filtrate. In this study we conducted bioassay with the fungal culture filtrate and culture fluid to greenhouse pests such as cotton aphid, whitefly and thrips. Culture fluid showed high mortality against green peach aphid, as well as cotton aphid, sweet potato whitefly, and western flower thrips. However, control effect of culture filtrate varied with pests. Culture filtrate showed high mortality to cotton aphid. Mortality of western flower thrips with culture filtrate was slower than culture fluid including spores and sweet potato whitefly was much lower. These results indicated that the culture fluid of Beauveria bassiana Bb08 has potential to simultaneously control various greenhouse pests.

Bemisia tabaci, sweetpotato whitefly, has been recognized one of the most destructive insect pests worldwide because of increased resistance to some insecticide groups requiring alternative strategies for its control. We conducted a study of the influence of relative humidity, temperature and different developmental stages on the susceptibility of sweetpotato whitefly to conidia of Isaria javanica isolate, which had been reported high virulence against Q biotype of B. tabaci. The mortality of tobacco whitefly was low at low constant relative humidities, but was high when kept high humidity for first 24 hours and transferred to low humidity. The Isaria isolate had wide range of temperature (15℃ to 35℃) to control sweetpotato whitefly. The isolate has virulence to the egg and all developmental stages of nymph of B. tabaci. These results indicated that the isolate had good control effects at various environmental conditions and is an excellent candidate to develop a microbial pesticide to control sweetpotato whitefly.

Sweetpotato whitefly (Bemisia tabaci), especially Q biotype, has been recognized one of the most destructive insect pests worldwide because of increased resistance to some insecticide groups requiring alternative strategies for its control. We studied the conidia production of entomopathogenic fungus Isaria javanica Pf04, which had been reported high virulence isolate against Q biotype of B. tabaci, using grain. Brown rice was most suitable for conidia mass production of the isolate of I. javanica. Conidia was produced high at 25 ~ 27.5℃. The isolate produced more spores when conidia suspension directly inoculated onto media than two-phase fermentation. When concentration of inoculum was high spore production was high, but increasing rate of conidia production was highest at low inoculum concentration (1×105 conidia/ml) as 6,700 times increase compared with 20 times increase at high inoculum concentration (1×108 conidia/ml). These results indicated that the isolate can produce more conidia with cheap agricultural product and can develop as a microbial pesticide to control sweetpotato whitefly.