Mealybugs (Hemiptera: Pseudococcidae) are major pests of a wide range of crops and ornamental plants worldwide, and represented by 56 species in Korea. Occurrence of, or damage by, the longtailed mealybug, Pseudococcus longispinus Targioni-Tozzetti has not been reported since 2002. However, for the first time in many years, P. longispinus was reported at Phalaenopsis orchids by Kim et al in 2015. Therefore, the present study aimed to investigate the distribution of P. longispinus which is usually found inside a greenhouse or a building where the host plants are. We have collected 129 mealybug samples at 254 sites from Korea. Among these, 27 of P. longispinus and 62 of an unidentified Pseudococcus species were found mainly in ornamental plants such as Heteropanax fragrans, commonly known as happy trees in Korea. Morphological characteristics of the unidentified mealybug were very similar to those of P. longispinus. However, the genetic variability in COI sequence comparison was significantly different between the two species. Further studies are needed to clarify the valid name for this unidentified species, and we are currently working on it.

Hypocreales entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and 004, and produced in three grains, as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions. The conidia of JEF 007 and 003 wild type and AtMT-based generating random mutants were subjected to SDS-PAGE. A significant relationship between conidial thermotoelrance and detected proteins was observed. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

Bean bug, Riptortus pedestris is an agriculturally serious pest in north eastern Asian countries, damaging to several legumes and fruit trees. Chemical pesticides have been largely used to control the pest but it encounters insecticide-resistance and environmental toxicity issues. Alternatively different mode of action and environmentally sound pest management system can be found in entomopathogenic fungal insecticides. Herein we developed a platform to optimize the fungal production to express their maximum virulence against bean bug, by focusing on solid culture system for thermotolerance, formulation to select effective surfactants to carry the fungal conidia on the cuticles, and relationship between environmental abiotic factors and fungal mortality. First to produce highly thermotolerance fungal conidia, Beauveria bassiana and Metarhizium anisopliae isolates were cultured on several granular cereal substrates, which could be subjected to formulation process. Among the tested media, four media (millet, non-glutinous italian foxtail millet, glutinous italian foxtail millet, brown rice) were superior to the other grains in the spore production and thermotolerance. Next to efficiently deliver the fungal conidia on the cuticles of bean bug, total of six surfactant (CO-2.5, CO-12, LE-7, PE-61, TED-3 and Siloxane) was used to experiment. CO-12 was superior to the other surfactant in mortality of 100 ppm consistence. This work suggests that solid culture system and formulation and application should be seriously considered to reach an optimal level of mortality by inducing their maximum virulence.

Entomopathogenic fungi are facultative microorganisms, dwelling in soil or infecting host insects, and some of the genera have been used as biological control agents worldwide. Collection of fungal isolates should be a platform for the development of highly effective resources, thus 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 three was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, spectrum of virulence, cultural characteristics, thermo-stability of fungi, 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.

ocust, Locusta migratoria (Orthoptera: Acrididae) is one of the outbreaking pests worldwide and such big occurrence was recorded in 2014, Korea, however little consideration was given to the management strategy of the pest. Herein we established a indoor locust-rearing system and constructed a locust-pathogenic fungal library to further facilitate the resources to be used as possible biological control agents. A locust colony was provided from the National Institute of Agricultural Science and Technology and reared in corn or barley plants at artificially manipulated rooms. The critical developmental stages, such as oviposition, hatching and mating were successfully proceeded. Entomopathogenic fungal granules were treated to the locust (2 g/rearing box), and in 5~7 days mycosis was observed in the membranous cuticles of head, abdomen and legs. In particular JEF-003 (Metarhizium anisopliae), JEF-186 (M. lepidiotae) and JEF-187 (Clonostachys rogersoniana) showed high virulence against the locust. A population of locust was exposed to the entomopathogenic fungal conidia-incorporated soil to investigate the possibility of the fungal isolation from natural soil, which resulted in the pathogenesis in 7~10 days in laboratory conditions. More than 80% of control efficacy was observed in the greenhouse trial of fungal granular application. This work suggests that locust rearing system was successfully established and entomopathogenic fungi can be used to control the migratory locust.

Entomopathogenic Beauveria bassiana and Metarhizium anisopliae are well-known biological control agents worldwide and have high potential in industrialization. However their thermo-susceptibility limits long-term storage under high temperature conditions and high insecticidal activity after application to target pests. Herein we isolated highly virulent isolates, B. bassiana JEF006 and JEF007 and M. anisopliae JEF003 and JEF004, and produced in three grains, such as sorghum, millet and Italian millet as substrates for solid cultures, followed by thermotolerance assays to compare the potential of the three substrates for thermotolerance. The JEF isolates were exposed to dry and wet heat at 50°C and overall conidia were more stable under dry heat condition rather than wet heat. Of the three grains, Italian millet was superior to the other grains in the production of thermotolerant conidia. Additionally Italian millet did not severely aggregated, which enabled air to penetrate into the substrate well compared to the sorghum and millet. JEF isolates were more thermotolerant when they were kept in oil conditions as carriers of an oil-based formulation. This work suggests that Italian millet can be used as an effective substrate to produce more thermotolerant conidia, thus maintaining viability for long times under unfavorable environment and biological activity against target pests.

다공성 전극표면에 이온교환고분자를 직접 casting하여 만들어진 복합탄소전극의 성능을 알아보기 위해서 NaCl 수용액을 이용하여 흡착시간, 공급액 농도, 유속, 탈착전압에 따라 흡/탈착실험을 진행하였다. 유입수가 100 mg/L일 때 동일 조건에서 흡착시간이 3분에서 5분으로 증가하면서 제거율이 3% 증가하였는데 이는 유입수의 셀 내부 잔류시간의 증가로 인 한 것으로 사료되며 또한 유속이 15 mL/min에서 23 mL/min 증가하면서 효율이 12% 정도 낮음을 보인 것은 유속이 상승하 면서 유입수의 셀 내부 잔류시간이 짧아지면서 나타나는 영향으로 사료된다. 유입수의 농도를 200 mg/L로 증가하였을 때 효 율은 100 mg/L보다 10~15% 정도 낮은 값을 보였는데, 이는 탈착구간에서 완전탈착이 되지 않아 나타나는 것으로 판단된다.

Mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae) has high and safe protein contents, which enables it to be animal feed. However, occurrence of entomopathogenic fungi in mealworms is one of the limitations for mass production. In this work, we investigated relationships between abiotic conditions and occurrence of fungal pathogens and established an effective control method using fungicides. In virulence assay, third instar mealworm larvae were sprayed by six entomopathogenic Beauveria bassiana isolates and kept under high relative humidity; B. bassiana ERL1575 isolate had highest virulence. Under normal humidity, ERL1575 conidia showed different virulence between spray (~0% virulence) and digestion (~80% virulence) method. Furthermore, mealworms, which digested conidia, were exposed to various temperature (20-35°C) and humidity (1-3 ml distilled water spray/35 mm diam. dish) conditions for 5 days. All the treatments showed ~90% virulence except 35°C incubations (~20% virulence), but irrespective to the humidity conditions. Forty chemical fungicides were assayed against conidial germination and hyphal growth of ERL1575. Fluazinam and mancozeb showed strong inhibition of conidial germination at standard application dose (SD), 1/2 SD and 1/5 SD; besides, fluazinam showed strong inhibition of hyphal growth. When fluazinam and mancozeb were applied to the fungal conidia-inoculated wheat bran, most of mealworms were alive after 3 days post application. However, high mortality rate (~100%) were observed in the conidia-inoculated wheat bran without any fungicides. In conclusion, this work suggests that B. bassiana isolates could be pathogens at <30°C when they were digested by mealworms, and fluazinam and mancozeb would be used as effective control agents against the pathogen.

고농도의 초산수용액을 분리⋅농축하기 위해 polyacrylonitrile (PAN) 중공사 복합막을 제조한 후 투과증발 분리실험을 통해 제조된 복합막의 분리성능을 확인하였다. 제조된 중공사 복합막의 코팅여부를 알아보고자 SEM 사진을 측정하였으며 코팅두께가 3.85 µm로 균일하게 코팅된 것을 확인할 수 있었다. 또한 막의 투과성능평가 실험을 수행한 결과 가교제의 농도와 가교온도가 증가함에 따라 투과도는 감소하고 선택도는 증가하는 경향을 나타내었고, 대표적인 결과로 가교제인 glutaraldehyde 농도 13 wt%, 가교온도 140°C에서 투과도 250 g/m2⋅hr, 선택도 13의 결과를 얻을 수 있었으며, 또 다른 가교제로서 poly (acrylic acid) 농도 9 wt%, 가교온도 140°C일 때 투과도 330 g/m2⋅hr, 선택도 9의 값을 얻을 수 있었다.

Background and Purpose: The Ankle-Brachial Index (ABI) is the ratio of blood pressure in the lower legs to that in the arms. The intima-media thickness (IMT) of extracranial carotid arteries determined by B-mode ultrasound is a measurable index of the presence of atherosclerosis. A low ABI and a high carotid IMT are independently related to increased risk of cardiovascular events. This study examined the association between carotid IMT and ABI in patients with ischemic stroke. Materials and Methods: Retrospectively, 116 hospitalized patients with ischemic stroke were recruited. Using a pulse wave velocity ABI device along with carotid duplex sonography, we measured carotid IMT and ABI and investigated the correlation between average values. Results: There was a significant difference in carotid IMT between the normal and abnormal ABI groups (P=0.0262). The group with an abnormal ABI was more than five times as likely to have increased carotid IMT as the group with a normal ABI (age, sex-adjusted OR 5.67 (95% CI 1.85~17.38)). The ABI and carotid IMT showed a weak inverse linear correlation in patients with ischemic stroke (correlation coefficient −0.378 after adjusting for age and sex). Conclusion: Our study suggests that an abnormal ABI is associated with a high carotid IMT in patients with ischemic stroke.

The legume family (Fabaceae) is the third largest family of flowering plants worldwide and often damaged by stink bugs, particularly Riptortus clavatus Thunberg (Heteroptera: Alydidae). This insect selectively feeds and oviposits on these plants, resulting in >50% of damage to soybeans harvested in autumn. The most common control methods for this insect are periodic application of chemical pesticides, pheromone traps, and bait crops, but still they encounters residual problem, insect resistance or low control efficacy. The current management system needs alternative control methods, which possibly combined with the present methods. Some entomopathogenic fungi have high virulence against the nymphs and adults of R. clavatus in a favorable conditions, reaching ∼80% mortality with mycosis in 3∼5 days. The genera of Beauveria and Metarhizium are possible control agents for effective and safe management of the serious pest. Fungal infection in R. clavatus was observed using an egfp-expressing fungal transformant. Thermotolerant entomopathogenic fungal conidia can be produced in cereal substrates, which enables conidia to be stable for long times. Our considerations need to be given to the combinations with other control methods, such as chemical pesticides or pheromone trap.

Antimicrobial peptides (AMPs) can be produced in mealworms. In this work, we integrated Bombyx mori (Bm) AMP, cecropin A to Beauveria bassiana ERL1170 by restriction enzyme-mediated integration method, which was confirmed by RT-PCR and an antibacterial activity assay. For the extracellular secretion of Bm cecropin A protein, the active domain of the cecropin A gene was tailed to the signal sequence of B. bassiana chitinase (Bbs). To exchange Bbs-cecropin A gene with egfp gene in pBARKS1-egfp, Bbs-cecropin A fragment was cut from pGEMBbs-cecropin A using XbaI/blunted and BamHI and ligated with cut pBARKS1-egfp using NcoI/blunted and BamHI, designated as pBARKS1-Bbs-cecropin A. After the transformation, transformants were grown on Czapek’s solution agar containing 600 μg ml-1PPT. Expression of Bm cecropin A was confirmed by RT-PCR. Strong clear zone was observed in the co-culture of the transformant D-6 and Bacillus subtilis on fourth strength Sabouraud dextrose agar 1 day after the culture at 25°C, whereas the wild type had no clear zone. This work suggests that Bm cecropin A can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.

Bean bug, Riptortus clavatus Thunberg (Heteroptera: Alydidae) causes serious damage to Leguminosae. Herein an entomopathogenic fungal virulence assay system against bean bugs was established to construct a fungal database which can be used in integrated pest management (IPM). First to obtain as many bean bugs as possible at the same stage, host plant-preference and developmental synchronization of bean bugs were investigated. In the preference assay, five pairs of adults were infested in a plastic cage, where a pot of green bean, pea or cowpea was previously placed. The highest fecundity and the fastest development of bean bug was observed in the green bean cage. Secondly, in the synchronization experiment, eggs were collected from the cage of adults in 1, 3, 5 and 7 days after oviposition and transferred to a fresh cage with green beans. From the every 4 days of survey, similar stages of bean bugs were found in the cages with the oviposition for 1 and 3 days, rather than the longer times of oviposition. A fungal bioassay against bean bugs was conducted using the bean bugs from the above insect rearing system. Ten Beauveria bassiana isolates were cultured on quarter-strength Sabouraud dextrose agar (¼SDA) for 7 days at 25°C. Ten 4th instar of nymphs were placed on a cultured plate for 1 hour and tranferred to a fresh moisturized plate with grains of green bean. ERL836 isolate treatment showed the highest virulence and fungal mycosis was observed on the bean bugs. In conclusion, these results can be useful to establish an entomopathogenic fungal database for IPM.

본 연구는 건조 공정을 통하여 인삼의 생리 활성을 유지하기 위해 수행되었다. 인삼의 생리활성을 유지하면서 인삼을 건조하기 위해서 낮은 온도와 압력에서의 균형 잡힌 감압 건조 공정을 적용하였다. 감압 건조 공정은 일반적인 열풍 건조보다 긴 공정 시간이 걸리지만 훨씬 낮은 온도에서 건조를 진행할 수 있기 때문에 열에 불안정한 인삼의 다당류의 파괴를 막을 수 있으며, 동결건조 공정보다 낮은 에너지로도 비슷한 생리 활성을 유지할 수 있어 경제적으로도 이점이 있다. 이러한 결과는 열풍건조 공정의 단점인 높은 온도로 인한 페놀성 화합물과 같은 휘발성 생리 활성 성분들의 파괴를 줄여 높은 항산화 활성을 유지하였다. 더 나아가, 감압건조 공정을 통한 건조인삼의 열수 추출물은 열풍건조와 동결건조 공정을 통한 건조인삼 보다 면역 B세포와 T 세포의 생육도가 증진되었으며, IL-6와 TNF-α와 같은 cytokine의 분비량도 증가하였다. 이러한 결과는 낮은 온도와 압력의 감압 건조 공정을 인삼에 함유된 진세노사이드 외의 열에 약한 성분들의 파괴를 막아 생리활성의 저하를 줄일 수 있을 것으로 사료된다.

Poly (vinylidene fluoride) (PVDF)ㆍpoly (acrylonitrile) (PAN) 중공사막에 친수성 고분자인 poly (vinyl alcohol) (PVA)과 가교제인 poly (acrylic acid) (PAA)을 코팅하여 복합막을 제조하였다. 제조된 막의 투과특성평가를 위해 90 wt% 물- 에탄올 혼합액에 대한 투과증발실험을 수행하였으며, 반응온도, PAA용액의 농도, 공급액의 온도변화에 따른 투과도 및 선택도를 측정하였다. 일반적으로 반응온도, PAA용액의 농도가 증가할수록 투과도는 낮아지고 선택도는 증가하는 경향을 보였으며, 공급액의 온도가 증가할수록 투과도는 증가하고 선택도는 낮아지는 경향을 보였다. 대표적으로 PVDF중공사 막은 투과도는 PAA 3 wt%, 반응온도 60°C, 공급액 온도 70°C에서 502 g/m 2 hr, 선택도는 PAA 11 wt%, 반응온도 100°C, 공급액 온도 50°C일 때, 218의 결과를 얻을 수 있었다.

Antimicrobial peptides (AMPs) can be produced in mealworms, currently being used as animal feeds, by the infection of genetically engineered-entomopathogenic fungi. In this work, we integrated Bombyx mori (Bm) AMP, cecropin A to Beauveria bassiana ERL1170 by restriction enzyme-mediated integration method, which was confirmed by RT-PCR and an antibacterial activity assay. For the extracellular secretion of Bm cecropin A protein, the active domain of the cecropin A gene was tailed to the signal sequence of B. bassiana chitinase (Bbs). To exchange Bbs-cecropin A gene with egfp gene in pBARKS1-egfp, Bbs-cecropin A fragment was cut from pGEM-Bbs-cecropin A using XbaI/blunted and BamHI and ligated with cut pBARKS1-egfp using NcoI/blunted and BamHI, designated as pBARKS1-Bbscecropin A. After the transformation, transformants were grown on Czapek’s solution agar containing 600 μg ml-1PPT. Expression of Bm cecropin A was confirmed by RT-PCR. Strong clear zone was observed in the co-culture of the transformant D-6 and Bacillus subtilis on fourth strength Sabouraud dextrose agar 1 day after the culture at 25°C, whereas the wild type had no clear zone. This work suggests that Bm cecropin A can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.

Entomopathogenic fungi formulated as wettable powders and suspension concentrates have been sprayed to crop pests for pest management. However, the use of fungal granules to control paddy field pests has not been fully explored. Herein, several Beauveria bassiana isolates (ERL 1170, 1578 and 836) were produced as granules using a millet-based solid culture. The granules were applied to the rice nursery 3 days before transplanting and their control efficacy against rice water weevils was determined in paddy fields. The solid cultures produced ~1×108conidiag-1ofmilletgrains10daysaftertheinoculation. The granules were applied to the soil in the rice nursery 3 days before the rice seedlings were transplanted in the paddy fields. Rice in plots with granules of ERL1578 had 17.3% leaf damage (74% control efficacy) 14 days post application, whereas rice plants in the non-treated control had 66.5% damage. Rice plants treated in the nursery with ERL1170 and ERL836 had 52~54% damage. In the rice plots previously treated with ERL1578 the smallest numbers of larvae and adults were observed 38 days post application. In laboratory conditions, ERL1578-treated larvae were tuned pink and covered with mycelial mass. Applications of millet-based B. bassiana granules on rice nursery soil can be an effective and efficient biological control strategy for the management of rice water weevils. This method is relatively inexpensive and requires less labor compared to practices involving the spraying of fungi directly on rice in paddy fields.

Brown planthopper, Nilaparvata lugens is giving enormous damage to rice production. In this work, virulence of four Beauveria bassiana isolates against brown planthoppers was investigated by applying fungal granules on the water of pots, and we further examined the growth of hypha on the rice plants from the water surcae to explain the insecticidal mode of action. We used Beauveria bassiana (Bb) ERL 836, 1170, 1575 and 1578 isolates, which produced ~2×108 conidia/g of millet grain in a solid culture. Rice seeding was grown in breeding boxes at 28±2℃ for 5 days. Mycotized millet grains were treated on the water in a box at 1 g/box and the rice seeding was infested with 18~25 brown planthopper adults per box. A chemical pesticide and water treatment served as controls. Among the treatments, Bb ERL 836-treated plants had the lowest damage, rather than the other fungal treatments in laboratory assays. Hyphal growth on the stem of rice plants was observed in Petridish conditions under a fluorescent microscopy. This work suggests a possible control of brown planthoppers using entomopathogenic fungi.

Poly vinylidene fluoride (PVDF) 중공사막을 polyethylenimine (PEI)와 p-xylylene dichloride (XDC)를 이용해 친 수화 시킨 후 poly(vinyl alcohol) (PVA)과 가교제인 poly(acrylic acid-co-maleic acid) (PAM) 혼합용액을 코팅하여 막을 제조 하였다. 중공사막 표면의 코팅여부는 scanning electron microscope (SEM)을 통해 관찰하였으며, 막의 특성평가를 위해 물/에 탄올 혼합액에 대한 투과증발 실험을 수행하였다. 공급액 조성은 90 wt% 에탄올 수용액을 사용하였으며, 가교제 농도, 공급 액과 반응온도 변화에 따른 투과도 및 선택도를 측정하였다. 투과도는 반응온도 100°C, PAM 농도 3 wt%, 공급액온도 70°C 에서 1,480 g/m 2 hr, 그리고 선택도는 반응온도 100°C, PAM 농도 15 wt%, 공급액온도 25°C에서 αW/E = 82의 결과를 얻을 수 있었다.