• Journal of Veterinary Science
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• 제21권1호
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• pp.4.1-4.9
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• 2020

Fast and accurate detection of viral RNA pathogens is important in apiculture. A polymerase chain reaction (PCR)-based detection method has been developed, which is simple, specific, and sensitive. In this study, we rapidly (in 1 min) synthesized cDNA from the RNA of deformed wing virus (DWV)-infected bees (Apis mellifera), and then, within 10 min, amplified the target cDNA by ultra-rapid qPCR. The PCR products were hybridized to a DNA-chip for confirmation of target gene specificity. The results of this study suggest that our method might be a useful tool for detecting DWV, as well as for the diagnosis of RNA virus-mediated diseases on-site.

• International Journal of Industrial Entomology and Biomaterials
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• 제13권2호
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• pp.57-61
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• 2006

In this study, morphology of perithecia, asci, ascospores, etc. of C. sphecocephala were examined for its telemorphic characteristics. Its colony grew up to 32 mm in diameter on potato dextrose agar (PDA) for 30 days under the condition of $24{\pm}1^{\circ}C$. PDBLA and PDBAA media were selected as optimal media for C. sphecocephala, on which the growth was 1.5 times as fast as on PDA medium. Moreover, PDBLA medium induced successfully the synnemata of anamorphic state. C. sphecocephala was able to be proliferated in vitro on both larva and adult of honeybee drone as its substrate. After inoculated onto the drone larva, it produced mycelium at $24{\pm}1^{\circ}C$, with the maximum yield up to $67{\pm}3mg$ on the $50^{th}$ day.

• 한국양봉학회지
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• 제34권2호
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• pp.137-140
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• 2019

Although honeybees (Apis mellifera) are crucial for maintenance of the ecosystem, population of honeybee has been steadily decreasing due to diseases including nosemosis. Nosemosis is a disease caused by Nosema ceranae and is now considered as a major threat to honeybees. N. ceranae is a microsporidian that stays in form of spore even before the infection, which makes it harder to control than other pathogens. People are now aware of this parasite, however, cure and preventive candidates for nosemosis are hardly found until today. In this study, in vitro experiment of Lespedeza cuneata treatment to prevent nosemosis were done using Trichoplusia ni cell line, BTI-TN5B1-4. Normal T. ni cells exhibited round shape without abnormal size. On the other hand, when N. ceranae were treated, cells deteriorated and some cells abnormally enlarged due to N. ceranae infection. Interestingly, treatment of T. ni cells with L. cuneate extract protected abnormal cell shape induced by N. ceranae infection to normal shape. Some N. ceranae spores were observed outside of the cells. Effective concentration range for N. ceranae control were experimented. Lowest concentration which can control nosemosis were 50 ㎍/mL. When the concentration of L. cuneata extract was exceeded 200 ㎍/mL, cytotoxicity started to show up.

• 한국양봉학회지
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• 제34권4호
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• pp.285-293
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• 2019

The honey bee queen shows extreme polyandry and controlling the mating partners can only be possible either by artificial insemination or having remote isolated mating locations. Here we report on the German North Sea island of Heligoland. Because of its location 60 km from the mainland, the lack of a local population of honey bees, its size of just 1.4 ㎢ and suitable weather conditions during the months of May to July, it is considered an ideal location for controlled inseminations of high-quality virgin queen bees with drones deemed genetically superior to others. Methods how to rear virgin queen bees are described and information is provided on the numbers of queen bees, their supporting workers and drone bees that are taken to the island in the mating season. The bee most commonly involved in the Heligoland mating trials has become Apis mellifera carnica strain "Baltica". In one summer, for example, 80 virgin queens (belonging to beekeepers from nine different locations in northern Germany) each with about 600 worker bees plus two drone populations of around 2,000 drones were taken by ship to Heligoland. On their return to the mainland no later than 3.5 weeks after the mating exercise, the beekeepers could register a mating success rate of 80%. This information can help operation management of the new remote mating centre of Weedo Island, Jeonbuk in Korea, which is currently under construction.

• 한국양봉학회지
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• 제34권2호
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• pp.131-136
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• 2019

Recently, number of honeybees (Apis mellifera) has visibly decreased because they are vulnerable to some diseases like American foulbrood disease. American foulbrood disease, which is caused by Paenibacillus larvae, is emerged as great cause of decrease in number of honeybees. After antibiotic-resistant strain emerged, it is now more difficult to treat those pathogens successfully. Researches on finding alternative antibacterial compound are ongoing. In this study, we examined the antibacterial effect of honokiol on P. larvae. Honokiol showed great antibacterial effect with minimum inhibitory concentration of 12.5 ㎍/mL and minimum bactericidal concentration of 50 ㎍/mL. An agar diffusion test also confirmed the anti-Paenibacillus larvae activity of honokiol with an inhibitory zone of 9±0.5 mm. Since honokiol is known to interact membrane of some bacteria, we measured 260 nm absorbing particles, which could be induced by leakage of cells, and confirmed that the leakage of P. larvae occurred in dose-dependent manners. However, result of crystal violet assay suggested that honokiol has only mild anti-biofilm formation effect on P. larvae, which means honokiol controls the bacteria by inducing the bursting of membrane. Finally, an additive effect of honokiol with tetracycline and terramycin was found using a checkerboard assay with a fractional inhibitory concentration index value of 0.5.

• 약학회지
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• 제56권4호
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• pp.254-259
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• 2012

To further access honeybee (Apis mellifera L.) venom (BV) as a cosmetic ingredient and potential external treatment for topical use, we investigated its ability to inhibit tyrosinase activity and melanin biosynthesis on melanogenesis in B16F1 melanoma cells. We found that BV increased the cell viability in B16F1 melanoma cell and BV (0.01~1 ${\mu}g/ml$) inhibited melanin synthesis in with 10 nM ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) for 48 h. In addition, we used reverse transcription-polymerase chain reaction and western blotting for me melanogenesis-related genes such as tyrosinase to examine the mechanisms underlying the inhibitory effects of BV on melanogensis. BV inhibited direct tyrosinase activity, which decreased melanin synthesis in ${\alpha}$-MSH stimulated B16F1 melanoma cells. Thease findings suggest that BV induces the downregulation of melanogenesis by inhibiting tyrosinase activation.

• 한국산림과학회지
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• 제106권4호
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• pp.509-517
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• 2017

본 연구는 소나무재선충병의 확산 저지를 위해서 티아클로프리드 약제를 지상연막방제로 살포하였을 때 잣나무림 내 비표적 곤충 군집에 미치는 영향을 평가하기 위해 수행되었다. 또한 티아클로프리드의 살충 효과를 확인하기 위해서 표적 곤충인 북방수염하늘소와 대표적인 방화 곤충인 꿀벌에 대한 영향을 조사하였다. 2016년 강원대학교 학술림 내 잣나무림에서 처리구간(대조구와 약량별 처리구) 곤충류의 개체수, 종수, 종 구성의 유사도 그리고 곤충 2종의 치사율을 비교하였다. 각 조사구에는 곤충류 조사를 위해 6개의 함정트랩과 2개의 다중깔때기트랩을 설치하였고, 연무연막 살포 후 치사된 곤충류 조사를 위해 3장의 방수포를 배치하였다. 또한 북방수염하늘소와 꿀벌을 망실에 넣어 조사구 중심부에 위치한 잣나무의 7 m와 15 m 높이에 걸어 두었다. 조사 결과, 티아클로프리드를 1회 살포한 본 연구의 경우에는 곤충 군집이나 비표적 곤충인 꿀벌에 대한 영향은 미미한 것으로 확인되었다. 반면에 표적 곤충인 북방수염하늘소의 경우에는 티아클로프리드에 의한 치사 효과가 확인되었다. 망실 내 북방수염하늘소의 치사율은 망실의 높이와 공간적인 위치에 따라 달랐는데, 이는 티아클로프리드의 연무연막이 임내의 미기상 영향을 많이 받기 때문인 것으로 판단된다. 따라서 연무연막기를 이용하여 잣나무림 내 북방수염하늘소의 밀도와 확산을 억제하기 위해서는 연막기술을 개선함으로써 살충 효과에 대한 불확실성을 최소화시킬 필요가 있을 것이다.

• 대한약침학회지
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• 제25권4호
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• pp.390-395
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• 2022

Objectives: Sweet bee venom (sBV) is purified from Apis mellifera, containing a high level of melittin-its main component. It has been used as a therapeutic agent for pain relief and anti-inflammation, as well as for treating neuronal abnormalities. Recently, there have been studies on the therapeutic application of sBV for anticancer treatment. In the present study, we investigated the pharmacological effect of sBV treatment in A549 human lung cancer cells. Methods: We used microscopic analysis to observe the morphological changes in A549 cells after sBV treatment. The MTT assay was used to examine the cytotoxic effect after dose-dependent sBV treatment. Molecular changes in sBV were evaluated by the expression of apoptosis marker proteins using western blot analysis. Results: Microscopic analysis suggested that the growth inhibitory effect occurred in a dose-dependent manner; however, cell lysis occurred at a concentration over 20 ㎍/mL of sBV. The MTT assay indicated that sBV treatment exhibited a growth inhibitory effect at a concentration over 5 ㎍/mL. On fluorescence activated cell sorting analysis, G0 dead cells were observed after G1 arrest at treatment concentrations up to 10 ㎍/mL. However, rapid cell rupture was observed at a concentration of 20 ㎍/mL. Western blot analysis demonstrated that sBV treatment modulated the expression of multiple cell death-related proteins, including cleaved-PARP, cleaved-caspase 9, p53, Bcl2, and Bax. Conclusion: sBV induced cell death in A549 human lung cancer cells at a pharmacological concentration, albeit causing hemolytic cell death at a high concentration.

• 한국식품과학회지
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• 제29권1호
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• pp.26-31
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• 1997

물을 극미량 함유하는 미수계 효소반응계인 aerosol-OT/이소옥탄으로 구성된 역미셀계내에서 레시친의 일종인 DPPC (dipalmitoyl phosphatidylcholine)의 가수분해반응을 phospholipase $A_2$가 촉매할 수 있었다. 또한 phospholipase $A_2$에 의한 가수분해반응을 정량분석하기 위한 고성능 액체 크로마토그래피에 의한 예민하고 간편한 방법을 고안하였는 바, 이에 의하여 aerosol-OT/이소옥탄 역미셀계내에서 phospholipase $A_2$에 의한 가수분해반응의 동력학적인 분석이 가능하였다. 한편, 여러가지 유화제와 유기용매 중 aerosol-OT와 이소옥탄일 때 효소반응이 가장 효과적이었으며, 반응 적정온도는 $35{\sim}40^{\circ}C$, 적정 pH는 7.0이었다. 또한 역미셀계내에서의 가수분해반응은 수분 함량변화에 민감하였으며, R-값(=[water]/[aerosol-OT])이 10.0일 때 가장 높은 효소활성을 나타내었다. 효소반응을 위한 최적조건하에서의 $K_{m,app.}$ 및 $V_{max.,app.}$는 각각 8.73 mM, 2.83 units/㎎ protein이었으며, 역미셀계내에서 phospholipase $A_2$에 의한 DPPC가수분해반응의 활성화에너지는 12.31 kcal/mole로 산출되었다.

• 한국응용곤충학회지
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• 제51권3호
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• pp.299-303
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• 2012

서양종 꿀벌 독의 채집시기와 지역에 따른 봉독의 성분 변화 및 약리효과에 미치는 영향을 검토하였다. 채집시기는 5월부터 9월까지, 채집지역은 전국 35개 지역으로부터 채취한 봉독을 대상으로 2010년과 2011년 2년에 걸쳐 동일 지역에서 동일한 방법으로 봉독을 채취하였다. 채취한 봉독은 액체크로마토그래피를 통해 멜리틴과 아파민 그리고 포스포리파아제 A2의 성분 함량을 분석하였다. 그 결과 채집시기와 지역에 따른 성분에 유의한 차이는 확인되지 않았다(One way-ANOVA, Duncan's test (${\alpha}$=0.05)). 봉독의 성분은 채집시기와 지역에 관계없이 멜리틴 $55.2{\pm}2.07%$, 아파민 $2.57{\pm}0.103%$ 그리고 포스포리파아제 A2는 $12.51{\pm}0.37%$을 차지하였다. 이상의 결과로부터 봉독은 채취시기에 따른 주요 성분은 차이를 갖고 있지 않았으며, 이는 꿀벌의 먹이, 사육온도 등 외부 환경이 봉독 분비에 영향을 주지 않는 것으로 사료되었다.