Thrips is one of the serious insect pests in many economic crops, in which general managements rely mainly on chemical insecticides. However, the overuse of chemicals led to the development of insect resistance and environmental residual issues. Therefore, thrips management requires the consideration of alternative strategies which are less harmful to the environment and working on different target points. In this work, we isolated entomopathogenic fungi from soil, and identified them with morphological and molecular biology methods, followed by a preliminary virulence assay against Tenebrio molitor larvae. Selected fungal isolates were adjusted to 1×107 conidia/ml for an indoor virulence assay against thrips adults, and highly virulent isolates were added to a thrip-pathogenic fungal library. Biological characteristics of efficacious isolates were investigated comparing to a previously commercialized fungal isolate. This entomopathogenic fungal library could be used as a valuable resource for developing effective strains to control thrips in agricultural fields.

Whitefly has a broad host range of more than 600 species, and 40 biotypes have been reported worldwide. Among the various ecological types, B and Q are the most problematic. Especially, Biotype Q, which causes serious damage in the green house, mediates more than 40 kinds of viruses, including Tomato yellow leaf cull virus (TYLCV). The development of resistance to whitefly, adverse effects on non-target organisms, and strengthened legal regulations are making chemical control difficult in application. For these reasons, the use of alternative strategies is needed to suppress the Biotype Q whitefly population. Entomopathogenic fungi were investigated and developed to control hard-control insect pest, such as whitefly. These fungi are natural pathogen of their invertebrate host and contribute to the regulation of their host population in the environment. In addition, these agents are typically harmless to humans and non-targeted organisms and do not adversely impact the environment. Therefore, in this study, we screened pathogenic fungi against whitefly using our entomopathogenic fungal library, and virulence tests were conducted with conoidal suspension (1×107conidia/ml) in laboratory conditions. As a result, 10 isolates showed 80-100% mortality at 5 days after treatment. We will analyze the characteristics of these fungal isolates and evaluate their potential as insecticides against whitefly. The selected isolates against the high mortality of whiteflies can be effectively used in integrated pest management programs for whiteflies.

Japanese pine sawyer beetle, Monochamus alternatus Hope (Coleoptera: Cerambycidae) is considered as a serious pest in pine trees. To develop an eco-friendly strategy to manage this forest insect, we collected entomopathogenic fungi from Korean soil and assessed their virulence against the adults of the insect in laboratory conditions. As a result, two isolates with conidial suspension (1.0×107conidia/ml), showed 87% and 90% mortality 12 days after fungal treatment, respectively. We assessed the potential of the fungi-derived destruxin and protease as additives to the fungal isolates, and they showed insecticidal activity via feeding and spraying treatments. Finally, we produced fungal conidia in massive solid cultures and formulated wettable powders, and now studying optimal conditions of oil-based formulation with two isolates. Based on these results, we are evaluating the control efficacy of the fungal agents against M. alternatus in field conditions.

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.

This study was aimed to evaluate the dose-response the effects of nano-encapsulated conjugated linoleic acids(CLAs) on in vitro ruminal fermentation profiles. A fistulated Holstein cow was used as a donor of rumen fluid. Nano-encapsulated CLAs(LF, 5% of nano-encapsulated CLA-FFA; HF, 10% of nano-encapsulated CLA-FFA; LT, 5% of nano-encapsulated CLA-TG; HT, 10% of nano-encapsulated CLA-TG) were added to the in vitro ruminal fermentation experiment. In the in vitro ruminal incubation test, the total gas production on incubation with nano-encapsulated CLAs was increased significantly according to the incubation time, compared with the control(p<0.05). The tVFA concentrations on addition of LF and HT were significantly higher than that of the control(p<0.05). Thus, nano-encapsulated CLAs might improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, the population of Butyrivibrio fibrisolvens which is closely related to ruminal biohydrogenation was increased by adding HT, while decreased by adding LF at 12 h incubation. These results showed that nano-encapsulated CLA-FFA could be applied to enhance CLA levels in ruminants by maintaining the stability of CLA without causing adverse effects on ruminal fermentation profiles considering the optimal dosage.

The fact that flip-flops, one of many different types of unstable shoes, are light and relatively easy to put on, accounts for their popularity among people. But because flip-flops rely heavily on the support of a single thong between your first and second toes, they impose a huge amount of pressure onto lower leg. Thus in the following experiment we tried to examine the different effects of flip-flops and running shoes in terms of their effect on muscle activity and fatigue of tibialis anterior and gastrocnemius during walking. In order to measure an electromyogram we used Free EMG system. 10 men and 10 women in running shoes ran on treadmills for 15 minutes at 4.8km/h, 2 days later the same experiment was carried out, but this time, in flip-flops. p value turned out to be greater than .05 and thus there was no considerable difference between the effects of flip-flops and running shoes on muscle activity and fatigue during walking. Therefore we conclude that despite the fact that flip-flops are considered unstable, their effects on muscle activity and fatigue of tibialis anterior and gastrocnemius are negligible.

Mealworms, Tenebrio molitor (L.) is used as an important animal feed additive for growth promotion and health management, but potentially exposes to fungal infection. In this work, virulence of two species of entomopathogenic fungi against the insect, and the relationship between abiotic features and virulence were investigated. Secondly our consideration was also given to the effect of chemical fungicides on conidial germination for risk control. Between Beauveria bassiana (Bb) and Metarhizium roberstii (Mr) (previously M. anisopliae), Bb isolates had much higher virulence (~100% mortality in 3~4 days after the treatment), rather than Mr isolates in laboratory assays. Next, fungus-treated mealworms were kept at wheat bran at 20, 25, 30 and 35℃ with 3, 6, 9 times of water spray to the feeds for set-up of different humidity conditions. Inoculation of fungi to mealworms was conducted by fungal spray and feeding methods, which resulted in higher virulence in feeding method. In the feeding method, all temperature treatments except 35℃ showed high virulence against mealworms, but any significant relationship between virulence and humidity was not observed. In the chemical fungicide screening, fluazinam (CAS No. 79622-59-6) and mancozeb (8018-01-7) significantly inhibited the germination of Bb and Mr conidia. This work suggest that contamination of wheat bran with fungal pathogens, particularly B. bassiana may induce mycosis of mealworms, but introduction of effective fungicides possibly reduce fungal infection.

LIVE/DEAD® BacLight™ and alamarBlue® are fluorescent materials used for the enumeration of live and dead bacteria. LIVE/DEAD® BacLight™ is generally used for confocal microscopy applications to differentiate live from dead bacteria in a biofilm or planktonic state. AlamarBlue® has also been used widely to assay live and dead bacteria in a planktonic state. Whilst these materials are successfully utilized in experiments to discriminate live from dead bacteria for several species of bacteria, the application of these techniques to oral bacteria is limited to the use of LIVE/DEAD® BacLight™ in biofilm studies. In our present study, we assessed whether these two methods could enumerate live and dead oral bacterial species in a planktonic state. We tested the reagents on Streptococcus mutans, Streptococcus sobrinus, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and Enterococcus faecalis and found that only LIVE/DEAD® BacLight™ could differentiate live from dead cells for all five of these oral strains. AlamarBlue® was not effective in this regard for P. gingivalis or A. actinomycetemcomitans. In addition, the differentiation of live and dead bacterial cells by alamarBlue® could not be performed for concentrations lower than 2 × 106 cells/ml. Our data thus indicate that LIVE/DEAD® BacLight™ is a more effective reagent for this analysis.

The cotton aphid, Aphis gossypii (Glover), is one of the most serious pest in seed potato and various vegetable cultivation. The imidacloprid-resistant strain (IR) was over 300-fold more resistant to imidacloprid compared to a susceptible strain (S) as judged by LC50 values. A highly imidacloprid-resistant local field population (L) was collected from cucumber at Gangwha island in 4th August 2011. Even though neonicotinoid insecticides especially imidacloprid were sprayed six times during June and July, aphid density was too high to be counted. To identify differentially expressed genes in IR or L, comparative transcriptome analyses based on GS-FLX were conducted using total RNAs extracted from IR, L and S strains. Futhermore, to search the resistance associated proteins in IR or L, comparative proteome analyses based on 2DE were conducted using total proteins extracted from IR, L and S strains. Few common candidate genes detected among IR and L such as ABC genes. Comparison of the nucleotide sequence of six nicotinic acetylcholine receptor (nAChR) subunit (alpha 1-5, beta 1) genes from IR, L and S strain revealed a point mutation in the loop D region of the nAChR beta 1 subunit of the IR, causing an arginine to threonine substitution (R81T). These mechanisms also reported in Myzus persicae and this amino acid change confers a vertebrate-like character to the insect nAChR and results in reduced sensitivity to neonicotinoids.

Historically the rise of contagious diseases and epidemics has been mainly caused by lack of food safety knowledge and negligence in personal hygiene. Food poisoning is an example that have been caused by lack of proper knowledge in food safety and personal hygiene. In order to prove this hypothesis, our team distributed a total of 225 survey sheets to middle school students. Survey questions mainly focused on the students' food safety knowledge, personal hygiene and regular dietary habits. We used 12 items to determine the stages of change in teenagers and divided into 3 stages of change: precontemplation stage (poor problem recognition), contemplation and preparation stage (some problem recognition), and action stage. Out of the 225 students surveyed, 105(47.1%) were male and 118 (52.9%) were female; 44.0% were in 7th grade, 33.0% in 8th grade, and 24.0% in 9th grade. In stages of change model, 3.8% were classified in pre-contemplation stage, 31.1% in preparation stage, and 65.1% in action stage. The awareness of the significance of food safety has a strong positive correlation with the stages of change food safety behavior (P<0.001), which suggesting our questionnaires regarding stages of change and food safety behavior were quite credential. Stages of change were also correlated with the consumption of unsanitary foods (P<0.01) and with the frequent hand washing (P<0.05). The stages of change showed a positive correlation with the food safety knowledge level (P<0.05). The four questions (out of 10) with the least percentage of correct answers were questions about the meaning of Norovirus (23.6%), the meaning of expiration date (43.6%), the meaning of food poisoning (36.4%), and proper food-handling (36.0%).

Ascotis selenaria, a major geometridae moth in citrus trees, annually damages the citrus leaves and fruits. The surface of young citrus fruit were usually fed by 1st larva of A. selenaria after landing or stepping onto the citrus fruits. To protect the larval damages of citrus fruits needs to predict the accurate occurrence time of the 1st larva for spraying. Because larval occurrences is dependent on the oviposition of adult female and the eggs were not found in/on citrus trees, oviposition model of A. selenaria linked with the egg development model will be helpful of protecting larval damages on citrus fruits. Adult longevity, survival and fecundity of A.selenaria was investigated at 13, 16, 20, 24, 28, 30, 32, and 35℃. The longevity decreased as the temperature increased and the female development rates (1/median longevity) were well described by a modified sigmoid model, which was used to calculate the adult physiological age. Description of the total fecundity was used by a non-linear model: The maximum fecundity of A. selenaria was estimated as 2490 eggs and peaked temperature was 19.7℃ according to the fecundity model. The cumulative age-specific oviposition rate and the age-specific survival rate was well described by 2 parameters Weibull function and a reverse logistic curve respectively. Total fecundity model, age-sepecific oviposition model, and age-specific survival model were incorporated into the oviposition model.

Oral spirochetes are anaerobes known as one of causative agents for periodontal diseases. In this study, we investigated the possibility of utilizing fluorescent fatty acids for labeling oral spirochetes. Bacterial labeling was standardized with three different lengths of fluorescent fatty acids: 5-octadecanoylaminfluorescein (OAF), 5-dodecanoylamin-fluorescein (DAF), and 5-hexadecanoylaminfluorescein (HAF). Among these fatty acids, OAF showed the best labeling activity. Treponema denticola ATCC 35405 was totally saturated to the maximum when incubated with OAF 1μM/mℓ for 1 hour. Treponema vincentii LA-1 also increased in fluorescence in proportion to incubation time length and the concentration. In conclusion, these findings showed the possibility that the fluorescent fatty acid can be used for labeling oral spirochetes.