비벡터링은 벌이 수정활동을 하는 동안에 이들로 하여금 병해충 방제용 미생물 제제를 식물체에 전달하게 하여 생력적으로 병해충 발생을 억제시킬 수 있게 하는 새로운 작물보호 기술이다. 국립농업과학원 시험포장과 농가포장의 방울토마토 시설하우스에서 화분매개충인 뒤영벌(Bombus terrestris), 미생물제제인 바실러스 서브틸리스(Bacillus subtilis), 그리고 자체 제작한 분배장치를 이용하여 비벡터링 시험을 수행하였고, 식물체에 전파된 미생물제제의 양과 비율을 측정하였다. 분배장치를 통과한 벌 충체에는 9.0x105∼1.9x106의 세균이 검출되었다. 농과원 시험포장 방울토마토 꽃에서 측정된 세균수는 꽃당 2,600-8,600개 였으며, 80-100%의 꽃에서 검출되었다. 미생물제제를 주 1회와 주 2회 교체한 경우 처리간에는 유의한 차이가 발견되지 않았다. 농가포장에서는 꽃당 1,800-2,400개를 83-93%의 꽃에서 검출할 수 있었고, 비벡터링시험전에 비해 세균수가 75배 증가되었다. 비벡터링 시험에 따른 벌 사망률은 22% 정도로 무처리구와 차이는 없었고, 따라서 미생물제제가 뒤영벌에 대한 부작용은 거의 없는 것으로 판단되었다. 비벡터링 기술 적용에 따른 방울토마토의 수확량은 무처리구와 비교해서 차이가 없었다. 앞으로 뒤영벌에 안전하면서 병해충에 대한 방제효과가 좋은 미생물 제제들을 선발하고, 병해충 발생에 따라 이 기술의 운용 전략 등을 세심하게 가다듬으면 화분매개곤충을이용하는 토마토, 딸기 재배에서 이 기술의 실용화는 매우 가까이에 왔다고 볼 수 있다.

Background: Propionibacterium acnes (P. acnes) is a major contributing factor for the inflammatory reaction of acne. Bee venom (BV) has been traditionally used to the treatment for inflammatory diseases. This study examined the anti-inflammatory effect of BV on P. acnes-induced inflammatory animal model. Methods: P. acnes were intradermally injected into both left and right ear of ICR mice. After injection, different concentrations of BV (1, 10 and 100 μg) mixed with 0.05 g of Vaseline was applied to the surface of the right ears of mice. Results: Histological observation revealed that P. acnes induced a considerable increase in the number of infiltrated inflammatory cells. However, BV treatment showed markedly reduced these reactions. Also, expression levels of TNF-α, and IL-1β were significant reduced in BV treated mice compared with P. acnes injected mice. The binding activity of NF-κB and AP-1 were increased in the P. acnes and Vaseline groups. In contrast, this enhancement of binding activity was markedly withdrawn after treatment with BV. Conclusion: In conclusion, this study indicates that BV has potential as an anti-acne agent and may be useful in the pharmaceutical and cosmetic industries.

Propionibacterium acnes (P. acnes) cause an inflammatory acne that plays an important role in the pathogenesis of acne by inducing inflammatory mediators. Bee venom therapy has been used in oriental medicine for the relief of pain and the treatment of inflammatory diseases. However, a direct effect of bee venom in skin inflammation has not been established. The purpose of this study was to investigate anti-inflammatory properties of bee venom in skin inflammation stimulated by heat-killed P. acnes using human keratinocytes and monocytes cell line. P. acnes stimulates the production of proinflammatory cytokines such as interleukins-1β, -8, interferon-γ and tumor necrosis factor-α in HaCaT and THP-1 cells. Bee venom effectively inhibits the secretion of IL-1β, IL-8, IFN-γ, and TNF-α. P. acnes treatment activates the expression of TLR2, which results in IL-8 expression. However, bee venom treatment reduces the expression of TLR2 and IL-8. Based on these results, bee venom has effects on anti-inflammatory activity against P. acnes in HaCaT and THP-1 cells.

For the investigation of the stability of purified bee venom(PBV) during the treatment in the temperature range of 50℃ to 120℃ for 24 hours, respectively, melittin contents, antibacterial effects, and cell regenerations were investigated. The changes in the melittin contents of PBV were not significantly different by treatment temperature below 70℃ for 24 houes and 80℃ for 4 hours. However the melttin contents is great decline after 24 hours above 80℃ for 24 hours. Antibacterial effects is not change below 80℃ for 4 houes but significantly decrease above 80℃ for 24 hours. Cell regenerations of PBV on human dermal fibroblast decreased at 80℃ for 24 houes, showing a significant difference from the below 80℃ for 4 houes. Through the temperature stability of PBV results of this study, it was treated that the melittin contents, antibacterial effects and cell regeneration effects of PBV could be maintained above 80℃ for 4 hours.

Sacbrood virus (SBV) is one of the most destructive honey bee virus. The virus causes failure to pupate and kills honey bee larvae. The infacted larvae`s color is change to brown. At the end, honey bee colony is destructed. Recently Korean Scabrood virus(KSBV) caused a great loss of Korean honey bee(Apis cerena) colonies for short period. Therefore, We need a highly rapid diagnosis method for rapid detection of KSBV. In this study, We need amicro-scale chip-based real-time PCR system (GeneChecker®). This system was developed for rapid, specific PCR based diagnosis. This system has uncommonly fast heating and cooling system. So We was able to detecting of KSBV in Apis cerena in short time. This system needs small reaction volume(total 10ul). This volume include SsoFast™ Evagreen Supermix and serially diluted cDNA templates showed a high sensitivity of 101copies.That machine can setting each PCR stage time. A specific detection primer set (KSBV-123-F/R) was used to amplify a unique 123bp DNA fragment. This PCR assays using serially diluted cDNA templates showed a high sensitivity of 101 copies. When applied to KSBV-positve samples, the result showed high specifity. The minimum diagnosis time was 9m 47s (30cycle). The amplied positive samples appear red fluorescent color. This novel detection method could be used a PCR-based diagnositic tool (GeneChecker®). The results showed high sensitivity and specifity in short time. And this diagnosis method is expected to be applied to rapidly detect various pathogens.

Black queen cell virus (BQCV), one of the most prevalent viruse, causes the death of queen larvae and pupae. The RNA-dependent RNA polymerases (RdRPs) are central components in the life cycle of RNA viruses that catalyzes the replication of RNA from an RNA template without DNA stage. Inhibition of RdRP gene is importantly significant for application of monoclonal antibody generation as a diagnosis tool for identifying BQCV infection in honey bee..In this study, the presence of BQCV in honey bee samples was confirmed by PCR using BQCV F/R primer set to multiply of 700 bp DNA fragment. For ampification of BQCV Rdrp gene, a primer set attached BamHI/SalI restriction site was designed based on the best homogenization between BQCV RdRP sequences in NCBI, a PCR product containing BQCV RdRP gene with 1576 bp in length was amplified. Furthermore, BQCV RdRP gene will be cloned into pBlueXcm vector for future researches.

We sequenced 17,329 bp of mitochondrial genome (mitogenome) of the black dwarf honey bee, Apis andreniformis (Hymenoptera: Apidae), that lacks ~200 bp of the A+T-rich region for the completion of the genomic sequence. The gene arrangement of A. andreniformis mitogenome is identical to that of A. cerana. However, the genome contains 5 additional tRNALeu(CUN) located 4 copies between tRNAMet and tRNAGln, and 1 copy between tRNAGln and tRNAAla, along with the typical sets of genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) including regular tRNALeu(CUN) and the A+T-rich region (at least 923 bp). Only 1 copy of tRNALeu(CUN) differed by 1 bp from other 4 copies of tRNALeu(CUN). Each additional tRNALeu(CUN) is followed by nearly identical 68-bp long repeat sequence (95.6% identity). All 13 protein coding genes have typical start codons found in insect mitochondrial PCGs (2 ATA, 9 ATT, and 2 ATG).

비벡터링(Bee-vectoring)은 벌통에 미생물제제가 담긴 분배장치를 내, 외부에 장착하여 벌들의 방화활동시에 미생물제제 전파활동을 동시에 수행하게끔 하는 기술로 국내에서는 최근들어 박 등(2009)에 의해 뒤영벌을 이용한 연구가 진행되 어오고 있다. 이 기술을 이용하여 효과적인 병해충 방제를 위해서는 뒤영벌에 안전 한 약제 선발이 중요한데, 본 연구에서 벌 활동수, 벌에 대한 사망률 결과를 토대로 암펠로마이세스 퀴스콸리스(Ampelomyces quisqualis) 94103 수화제, 바실루스 서 브틸리스 (Bacillus subtilis) KBC1010 수화제 2종을 새롭게 선발하였다. 또한 벌통 입구에 설치하는 분배장치를 개선하였다. 기존에 선보였던 장치와 비교하면 벌통 아래에서 고정되는 형태에서 벌통 위에 고정될 수 있도록 하였고, 분배장치의 출입 구를 전방쪽에서 측면방향으로, 이원화된 출입구가 하나가 되도록 개선하였다. 바 실루스 서브틸리스(B. subtilis) Y1336을 이용하여 2013년 봄 작기에 완숙 토마토 재배농가에서 실증한 결과, 미생물제제 매개량이 높았고, 잿빛곰팡이병 병발생률이 낮아짐을 확인할 수 있어 이 기술의 현장 적용가능성이 높음을 확인 할 수 있었다.

With over 7 billion people on the planet, agriculture faces immense pressure to meet global demands for food. One third of consumed food relies on insect pollination with by far, the predominate pollinator being the honey bee, Apis mellifera. Although future challenges facing agriculture will come from multiple domains, one of the immediate challenges is honey bee decline. Stress associated with transportation, pesticide exposure nutritional limitations, various diseases and pests have all been recognized as potential factors in honey bee decline. With the prospect of future global changes in climate, honey bees will also face changes in forage availability and overwintering potential. At the level of the individual colony, research has shown that honey bee health is directly correlated to genetic diversity. Increased colony diversity is associated with lower disease intensity, increased disease resistance, greater workforce productivity and thermoregulation stability. Genetic diversity at the population level serves as the raw material for selective breeding in agriculturally important plants and animals, including the honey bee. Honey bees are not native to Korea, however, and importation and founder events associated with the establishment of honey bees represent a series of genetic bottlenecks that limits the diversity of introduced honey bee populations. Fortunately, Apis mellifera consists of around 28 recognized subspecies within its native range, each with specific adaptations to climatic selective pressures endemic to its own location. Climate change is expected to be bring a high degree of uncertainty in the future to climate expression in various locations. Fortunately, the honey bee has a wide breadth of diversity contained within various subspecies and careful importation and evaluation of specific stocks may be highly useful as we enter climate uncertainty in the future. With the recognition that agro-ecosystems are highly interconnected and multifaceted, one of the greatest challenges facing agriculture is preserving and improving honey bee health.

In the present study, the 17,694-bp long complete mitochondrial genome (mitogenome) of the dwarf honey bee, Apis florea (Hymenoptera: Apidae), is described with an emphasis on the noteworthy triplicated tRNAser(AGN) region and an extraordinary long A+T-rich region with repeat regions. The gene arrangement of A. florea mitogenome is identical to that of A. mellifera, but has triplicated tRNASer(AGN), each of which contains the precedent 44 bp-long and following another 64 bp-long repeats plus one complete first repeat abutting to tRNAMet. A total of 1,610-bp long two repeat regions in 1,987 bp-long A+T-rich region is composed of nearly identical 141 ~ 219-bp long five tandem repeats and 50 ~ 52-bp long 12 tandem repeats that are encompassed by three non-repeat sequences. One of the common interpretations for such repeat sequence is slipped-strand mispairing and unequal crossing-over events during DNA replication.

Worldwide studies on Apis cerana variation for biogeography and genetic diversity depended largely on a 86~93 bp-long mitochondrial non-coding region (internal spacer region) located between tRNALeu and COII (named as NC2), possibly due to higher variability among available markers. In order to incorporate the A. cerana occurring in South Korea into world extensive data, we also sequenced the NC2 from 118 A. cerana samples collected over nine Korean localities and 66 A. cerana samples over seven Asian localities, such as China, Vietnam, and Thailand. These data were combined with preexisting world data to scrutinize genetic relationships of A. cerana in South Korea to outside distributional range. Sequencing of 184 samples provided a total of ten haplotypes: five from Korea, six from China, one from Vietnam, and two from Thailand. Among them eight were new, whereas two were previously reported ones. Phylogenetic analysis of A. cerana NC2 haplotypes so far found including ours has confirmed the presence of four major groups of A. cerana (Asian mainland group, Sundaland group, Palawan group, and Luzon-Mindahnao group) and all haplotypes found in this study also were included in the Asian mainland group. In order to find further variable regions that can be used as sequence-based marker several mitochondrial non-coding regions and nuclear intron regions are in the middle of testing.

Osteoarthritis is one of the commonest causes associated with age-related damage of articular cartilage. Non-steroidal anti-inflammatory drugs are commonly used in osteoarthritic patient. However, long term administration of these drugs results gastrointestinal disorders. Though, most studies have demonstrated in the past that bee venom has therapeutic effect on diseases related to inflammation and pains, but its anti-inflammatory properties have not been so far studied on inflamed chondrocytes (LPS induced) invitro. For the purpose, the study was carried out to determine the effect of bee venom on porcine articular chondrocyte cell using microarray. In this study, we found that 2,235 significantly associated gene (1,404 up-regulated genes and 831 down-regulated genes) that were expressed on inflamed and non inflamed chondrocytes during proliferation. Among the 1,404 up-regulated genes and 831 down-regulated genes, known genes were 372 and 237, respectively. On the other hand, bee venom significantly reduced expression of fetuin involved in acute inflammatory reaction. Our results suggest that this study could be useful database in gene expression profiling of chondrocyte cell treated with bee venom.

Nosema disease caused by Nosema apis which belongs to fungi is a major cause of honey production loss and is worldwide in distribution. N. apis infects the epithelial cells of the digestive system of adult honeybees. Nosema causes significant losses in population size of honeybee. There are about 25 thousand beekeepers caring for approximately 1,697,000 colonies in Korea. Honey production totaled almost 38,505 metric tons in 2010. This production was estimated to be worth about 274 billion Korean won. To determine infection level of nosema disease during the season, adult worker bees were collected from two colonies of experiment apiary from January to October. Our results indicate that the infection level of nosema disease was increased in spring and autumn. Also we initiated a survey of honeybee colonies on the blooming period of Acacia to determine the prevalence of N. apis. Twenty two hives owned by 18 beekeepers were sampled for this study. Bees were collected on 24th and 25th May of 2012. Nosema spore counts ranged from zero to 5,266,000 spores per bee. The average number of nosema spores per bee was calculated to be 1,375,000. Approximately 86% of the apiaries examined were infected with nosema, based on the presence of spores in the flowering period of Acacia. This indicates that nosema is the predominant species affecting honeybee colonies.

Bee-vectoring 기술은 벌이 작물에서 방화활동을 하는 동안 미생물제제를 식물체의 꽃이나 잎 등에 전파함으로써 병해충 방제를 가능케 하는 새로운 방제기술로 매개곤충, 미생물제제, 분배장치가 중요한 기술요소가 된다. 국내에서는 박 등(2009)에 의해 소개되었고, 지난 몇 년간의 연구에서 벌의 활동량, 제제 매개량을 높일 수 있는 분배 장치 등의 개발이 이루어져 왔다. 그러나, 수정벌이 안전하게 매개할 수 있는 미생물제제의 선발은 아직까지 이루어지지 않았다. 본 연구에서는 상업화된 미생물제제 제품중에서 이용 가능한 미생물제제를 선발하고자, 소형 케이지내에 농과원에서 개발한 분배장치를 뒤영벌 벌통에 연결하고, 벌이 분배장치를 통해 미생물제제를 묻혀나갈 때 벌에 묻혀나가는 양, 소형케이지내 벌의 활동성, 벌에 대한 안전성 등을 측정하였다. 벌의 사망률이 낮고, 벌의 활동에 부정적인 영향을 주지 않는 제제는 Bacillus thuringiensis kurstaki, Bacillus subtilis QST713, Bacillus subtilis Y1336, Simplicillium lamelicola BCP 였다. 또한 Bacillus subtilis QST713을 농가포장에서 검정한 결과, 소형케이지 시험과 일치되는 결과를 얻어 케이지시험이 미생물제제를 선발하는데 유용하다는 것을 확인할 수 있었다.

We investigated the molecular and kinetic properties of two acetylcholinesterases (AmAChE1 and AmAChE2) from the Western honey bee, Apis mellifera. Western blot analysis revealed that AmAChE2 has most of catalytic activity rather than AmAChE1, further suggesting that AmAChE2 is responsible for synaptic transmission in A. mellifera, in contrast to most other insects. AmAChE2 was predominately expressed in the ganglia and head containing the central nervous system (CNS), while AmAChE1 was abundantly observed not only in the CNS but also in the peripheral nervous system/non-neuronal tissues. Both AmAChEs exist as homodimers; the monomers are covalently connected via a disulfide bond under native conditions. However, AmAChE2 was associated with the cell membrane via the glycophosphatidylinositol anchor, while AmAChE1 was present as a soluble form. The two AmAChEs were functionally expressed with a baculovirus system. Kinetic analysis revealed that AmAChE2 has approximately 2,500-fold greater catalytic efficiency toward acetylthiocholine and butyrylthiocholine than AmAChE1, supporting the synaptic function of AmAChE2. In addition, AmAChE2 likely serves as the main target of the organophosphate (OP) and carbamate (CB) insecticides as judged by the lower IC50 values against AmAChE2 than against AmAChE1. When OP and CB insecticides were pre-incubated with a mixture of AmAChE1 and AmAChE2, asignificant reduction in the inhibition of AmAChE2 was observed, suggesting a protective role of AmAChE1 against xenobiotics. Taken together, based on their tissue distribution pattern, molecular and kinetic properties, AmAChE2 plays a major role in synaptic transmission, while AmAChE1 has non-neuronal functions, including chemical defense.

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 (=0.05)). 봉독의 성분은 채집시기와 지역에 관계없이 멜리틴 , 아파민 그리고 포스포리파아제 A2는 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.

The average life expectancy has continuously increased with the development of medicine. However, Some of men have been suffered from the insufficient secretion of testosterone causing by physical factors, social and psychological factors. Testosterone is an essential steroid hormone controlling male reproductive function. Alternative medicines in plants, fungi, and insects have been studied to enhance sexuality. In the present study, we determined the effect of an acorn pollen and Cordyceps militaris fruiting body on testosterone concentration in Sprague-Dawley rats. Eighteen rats per group were housed to regular diet or diet supplemented with acorn pollen powder or fruiting body power for 4 weeks. Serum was collected from 6 rats per group. Results showed that changes of the body weight, food and water intake of the rats were not observed in this study. However, acorn pollen powder and Cordyceps militaris fruiting body significantly stimulated testosterone production (p < 0.05) compared to the control, respectively. Hence, it suggests that acorn pollen or Cordyceps militaris fruiting body might be developed as a complementary medicine to improve sexual hormones.

This study was conducted in order to examine the safety of bee venom as an alternative for antibiotics using male ICR mice. Five-week-old male mice received a single intravenous injection of a dried honey bee venom at the concentration of 0.25 mg/kg (a clinical dose) or 0.5 mg/kg through the tail vein and various pathophysiological analyses were performed after three days. No significant differences in changes of body weight were observed between the saline-treated control group and the experimental groups. In the hematological analysis, none of the parameters were affected by bee venom. In blood biochemistry analysis, none of the markers were affected by administration of bee venom. Similarly, there were no significant effects on markers for liver, kidney, and skeletal muscle functions in all treated- groups. On macroscopic examination, no remarkable lesions were detected in these organs. Because there were no adverse effects of the bee venom in a single intravenous toxicity test for three days, it was concluded that bee venom could be a candidate for a safe natural antibiotic for use in the animal production industry.

Experiments were conducted in order to assess the healing effect of bee venom (BV) cream on full-thickness skin wounds in rabbits. BV cream was compared with silver sulfadiazine (SS) as a topical medicament against a control on experimentally created full-thickness wounds. Two wounds measuring 2 × 2 cm were created bilaterally (four wounds/rabbit) on the dorsolateral aspect of the trunk of seven New Zealand white rabbits. Wound treatments were evenly distributed on four sites, using a Latin square design. The contact layer of wounds was treated with physiological saline (control), SS cream, and BV cream over a period of 28 days. Assessment of wound healing was based on scab hardness, wound exudates, wound area, unepithelialized granulation tissue, and histopathological findings. Topical application of BV and SS creams to wounds resulted in reduced inflammation, debridement of necrotic tissue, and promoted granulation and epithelialization. Wound healing was faster, with statistical significance in BV and SS treatments, compared to the control (P<0.05). Treatment with BV evoked an anti-inflammation effect in a rabbit model. BV cream produced a wound healing effect similar to that of commercially available SS cream. Anti-inflammation effect as a topical treatment with BV cream appears to be better than that with SS cream. These results suggest that topical application of BV cream may be an alternative treatment for full-thickness skin wounds.

We present evidence that the serine protease found in bumblebee (Bombus terrestris) venom exhibits fibrin(ogen)olytic activity. Compared to honeybee (Apis mellifera) venom, bumblebee venom contains a higher content of serine protease, which is one of its major components. Venom serine proteases from bumblebees did not cross-react with antibodies against the honeybee venom serine protease. We provide functional evidence indicating that B. terrestris venom serine protease (Bt-VSP) acts as a fibrin(ogen)olytic enzyme. Bt-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products. However, Bt-VSP is not a plasminogen activator, and its fibrinolytic activity is less than that of plasmin. Taken together, our results define roles for Bt-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease, providing significant support for thepotential use of bumblebee venom serine protease as a clinical agent.