미포장충류(Nosema spp. (NS))는 양봉꿀벌에 심각한 문제를 야기시키는 기생충으로 효과적인 방제물질의 선발이 무엇보다 중요하다. 본 연구는 노제마병과 기타 꿀벌의 발생유행시기의 구명과 더불어 3가지 노제마병 방제물질(M1 = 벌꿀희석의 레몬쥬스; M2 = 설탕시럽 혼합의 카모마일 추출물; M3 = 설탕시럽 혼합의 항생물질 스트리베트)을 평가하고자 수행하였다. 꿀벌 성충과 유충집단의 질병 유행시기를 년간 조사하였으며, 야외 및 실험실 조건에서 노제마병에 대한 M1, M2, M3의 효과를 평가하였다. 조사결과 극소수의 꿀벌 성충과 유충 질병이 발견되었다. 노제마 병은 겨울과 봄 기간 저온과 고습조건에서 검출되었다. 포장실험에서 M2는 36.66%까지 발병억제 능력을 보였으며, 반면M3는 23.33%, M1는 13.33%의 억제효과를 보였다. 실내실험에서 M2가 방제효과가 가장 좋았고, 그 다음 M1와 M3 이었다. 3가지 방제물질은 병에 감염된 꿀벌성충의 생존력을 크게 높이는 것으로 나타났다. 본 연구는 노제마병 방제를 위한 천연물질로 카모마일의 잠재적 방제효과를 제시하고 있다.

The honey bee, Apis mellifera ligustica (Hymenoptera: Apidae), strain with a high hygienic behavior (HHB) has been bred for several years in Korea, and a diagnosis system to distinguish it from low hygienic behavior (LHB) strain has been necessitated. Thus, complete mitogenome of the two strains were sequenced through Next-Generation Sequencing technique to detect SNPs. Comparison of the mitogenome sequences from the two strains of A. m. ligustica have detected 23 SNPs in 11 PCGs and these were further confirmed the presence of SNPs using each 10 individuals selected randomly from each strain, indicating that these SNP markers might be useful to diagnose the honeybee strains with the HHB. Therefore, mitogenome sequences are promising genome source to detect SNP markers, particularly for inbred female iso-lines.

Honey bee has been widely used as a model insect for biological sciences because of its sociality and specialized labor division. For the investigation of the seasonal and labor-dependent expression patterns of genes putatively involved in its sociality, quantitative real-time PCR (qRT-PCR) can be applied to quantify gene expression level and selection of reliable reference gene(s) for normalization is an accurate step. In this study, using three softwares (geNorm, NormFinder and BestKeeper), we evaluated seasonal expression stabilities of four reference genes that have been widely used for qRT-PCR in forager and nurse heads. Among four candidates, two genes, rpS18 and gapdh, were suggested to be the optimal reference genes for qRT-PCR.

Honey bees are challenged with declining colony numbers, generally called ‘Colony Collapse Syndrome’ (CCS). This issue is certainly a syndrome because it is due to a range of threats, including parasitic mites, exposure to agricultural chemicals and various viruses, among other things. We posed the hypothesis the CCS could also be due to declining nutritional qualities, which may relate to digestive physiology. Because there is no information on honey bee lipid nutrition and digestion, we characterized a digestive phospholipase A2 in honey bee midguts. In this paper, I will report on some of our findings. With a focus on students, I will also use this report to share thoughts on presenting and writing for international audiences.

In this study, we investigated the nutritional composition including proximate, amino acid, vitamin, minerals, and the antioxidant activity of green tea (Camellia sinensis) pollen grains collected by Apis mellifera bees, for use as a health food. The crude protein and fat content was estimated at 26.14% and 3.49%, respectively. Eighteen amino acids were identified in green tea pollen, including 8 essential amino acids and 10 non-essential amino acids. The predominant amino acids were glutamic acid, proline and aspartic acid accounting for about 33.3% of total free amino acids. The concentration of vitamin C was the highest value of 35.7%, followed by B3 and B2 among the detected vitamins. The predominant minerals were potassium (790.32 mg/100g), followed by phosphorus (707.52 mg/100g) and sulfur (302.67 mg/100g), whereas copper, zinc and sodium were detected as minor elements. The antioxidant activity and phenolic content accounted for 33.8% at 500 μg/mL extract and 2.55 μg/mg, respectively.

Data clustering is one of the most difficult and challenging problems and can be formally considered as a particular kind of NP-hard grouping problems. The K-means algorithm is one of the most popular and widely used clustering method because it is easy to implement and very efficient. However, it has high possibility to trap in local optimum and high variation of solutions with different initials for the large data set. Therefore, we need study efficient computational intelligence method to find the global optimal solution in data clustering problem within limited computational time. The objective of this paper is to propose a combined artificial bee colony (CABC) with K-means for initialization and finalization to find optimal solution that is effective on data clustering optimization problem. The artificial bee colony (ABC) is an algorithm motivated by the intelligent behavior exhibited by honeybees when searching for food. The performance of ABC is better than or similar to other population-based algorithms with the added advantage of employing fewer control parameters. Our proposed CABC method is able to provide near optimal solution within reasonable time to balance the converged and diversified searches. In this paper, the experiment and analysis of clustering problems demonstrate that CABC is a competitive approach comparing to previous partitioning approaches in satisfactory results with respect to solution quality. We validate the performance of CABC using Iris, Wine, Glass, Vowel, and Cloud UCI machine learning repository datasets comparing to previous studies by experiment and analysis. Our proposed KABCK (K-means+ABC+K-means) is better than ABCK (ABC+K-means), KABC (K-means+ABC), ABC, and K-means in our simulations.

Among two different acetylcholinesterase (AmAChE1 and AmAChE2) of the western honey bee, the soluble AmAChE1might be related with a stress response as judged from its over-expression in honey bee workers when brood rearingwas suppressed. In this study, to ensure the nature of AmAChE1 responding to stress factors, the expression patternsof AmAChE1 were investigated following various treatments, including varroa mite infestation, bacterial challenge, broodrearing suppression, thermal stresses, chemical treatments, ultraviolet B irradiation, starvation, water restriction and crowdingstress. In addition, transcription profiles of four heat shock protein genes known as general stress markers and vitellogeningene, which is induced in several stress conditions, were tested as positive references. In every tested condition, onlybrood rearing suppression and heat shock were related with the expression of AmAChE1.

Bee venom contains a variety of toxic components, including vitellogenin, which display various biological, toxicological,and pharmacological activities. However, the biological actions of vitellogenin, a venom protein in bee venom, remainlargely unknown. Here, we demonstrate that Asiatic honeybee (Apis cerana) venom vitellogenin (AcV-Vg) exhibits anti-oxidantand anti-microbial activities. AcV-Vg is expressed in the venom gland and is then secreted into venom. The recombinantAcV-Vg protein was produced in baculovirus-infected insect cells. We found that AcV-Vg reduced cytotoxicity and oxidativedamage against oxidative stress. Furthermore, AcV-Vg bound to microbial surfaces and induced structural damage in themicrobial cell walls, which, in turn, exhibited anti-microbial activity against bacteria and fungi. Together, our data demonstratedthat the bee venom protein AcV-Vg has multifunctional roles as an anti-oxidant and anti-microbial agent.

서양종 꿀벌의 일벌의 독을 채취하여 정제한 정제봉독에 대한 세균에서의 돌연변이 유발성 검색을 위하여 S. typhimurium의 히스티딘 요구성 균주 TA100, TA1535, TA98, 그리고 TA1537의 균주와 대장균 E. coli의 트립토 판 요구성 균주인 WP2 uvrA를 이용해 복귀돌연변이시험 을 실시하였다. 정제봉독은 부형제로 사용한 멸균생리식 염수에 용해되었으며 모든 농도군의 조제물에서 침전은 관찰되지 않았으며, 정제봉독 조제물을 top agar와 혼합할 때 모든 농도군에서 혼탁이나 침전이 관찰되지 않았다. 대사활성계 적용 TA100, TA1535, TA98 균주에 대해 0, 1.5, 5, 15, 50, 150 및 500 μg/plate의 범위를 설정하고 미 적용시엔 0, 0.15, 0.5, 1.5, 5, 15 및 50 μg/plate의 범위로 설 정하였다. TA1537과 WP2 uvrA균주에 대한 농도 범위은 대사활성계 적용시엔 0, 5, 15, 50, 150, 500 및 1,500를 미 적용시엔 0, 1.5, 5, 15, 50, 150 및 500 μg/plate 범위로 설정하여 시험을 수행하였다. 그 결과 모든 시험균주에서 대사활성계 적용 여부에 상관없이 정제봉독 처리군 의 평균 집락 수는 증가를 나타내지 않았으며, 양성판정 기준을 만족시키지 못하였다. 따라서, 정제봉독은 본 시험 조건 하에 사용한 시험 균주에 복귀돌연변이를 유발하지 않는 것으로 사료되었다.

The nesting behavior, reproduction, fruit set and shape of O. cornifrons varied significantly with the released sex ratio of O. cornifrons. A female : male sex ratio of 1 : 2 was resulted in a 3.4 to 6.7 fold higher than other sex ratio in a nesting behavior. A ratio of 1 : 2 resulted in a 1.2-fold nesting rate, which was slightly higher than other nesting rates. Releasing only males resulted in a 2.4-fold greater amount of fruit set in non-pollinated sites. A sex ratio of 1 : 2 gave a slightly higher shape index and a 1.2 to 1.6-fold lower asymmetric index than other sex ratios. There was no significant difference between female release numbers in fruit set, and 100 to 200 females gave a slightly higher shape index than 400 females. Thus, we determined that 200 females should be released per 2,000㎡ and that the sex ratio of females to males should be 1 : 2.

Honey bee swarming is a natural phenomenon that occurs by changes of colony (i.e. population size and queen condition) and environment conditions. As cuticular hydrocarbons (CHCs) are known to be involved in the communication between honey bee nest-mates, we investigated and compared the CHC profiles of worker bees from individual colonies of 9-days before swarming (PPSC), a day before swarming (PSC), swarming (SG) and remaining (non-swarming) (RG). A total of 53 CHCs were identified by GC-MS, among which 11 compounds showed significantly differential expression patterns between swarming states. Before swarming (between PPSC and PSC), detection levels of 4 CHCs were significantly different, suggesting that production of some CHCs changed prior to swarming for swarming preparation. Six CHCs were deferentially produced between PSC and RG. The differential profiles of CHCs with respect to different swarming states are currently under investigation.

Perturbation of normal behaviors (e.g., nursing and foraging) in honey bee colonies by any external factors would immediately reduce the colony’s capacity for brood rearing, which can eventually lead to collapse of entire colony. To investigate the effects of brood rearing suppression in the biology of honey bee workers (nurse and forager), the gene-set enrichment analysis (GSEA) was performed for the transcriptomes of worker bees with or without their brood rearing being suppressed, from which functional profiles of pathways under influences by each condition were identified. Blocking of normal labor (i.e., nursing or foraging) induced the over-representation of pathways related with reshaping of worker bee physiology, suggesting that transition of labor is physiologically reversible. In addition, brood rearing suppression appeared to result in the reduction of neuronal excitability and aggressiveness in both forager and nurse, which would be necessary to manage the in-hive stress under unfavorable conditions

There are two different types of acetylcholinesterase (AChE1 and AChE2) in the western honeybee as in most of insects. It is suggested that soluble AmAChE1 might be related with a stress response as judged from its elevated expression level in honey bee workers when brood rearing was suppressed. In this study, to ensure the nature of AmAChE1 responding to stress factors, the expression patterns of AmAChE1 following heat shock, brood rearing suppression and chemical treatments (Imidacloprid and fluvalinate) were investigated. Also, several heat shock protein (hsp) genes (hsp10, hsp60, hsp70 and hsp90) known as general stress markers were tested as positive references. Heat shock induced expression of every tested hsp along with AmAChE1. In brood rearing-suppressed worker bees, 7 days old bees showed much higher expression level of AmAChE1 and hsp90 compared to control honey bees. However, treatment of imidacloprid and fluvalinate did not induce any apparent overexpression of these genes. These results confirm that both HSP and AmAChE1 genes generally respond to temperature and brood rearing suppression and further suggest that AmAChE1 can serve as a potential biomarker along with hsps for the detection of stress in honey bee colonies.

Honey production from approximately 1.7 million colonies owned by around 21 thousand Korean beekeepers was almost 36 thousand M/T in 2014. Honey farmers have mainly obtained commodities such as honey, beeswax, pollen and royal jelly. Honeybees gather the nectars and pollens from flowers of plants or trees. Bee pollen is rich in various nutrients and called as natural complete food. Bee pollens of Quercus acutissima, Actinidia arguta, Amorpha fruticosa, Oryza sativa, Chrysanthemum morifolium and Salix koreensis were mainly collected by Apis mellifera in Korea. But stiff pollen wall hinders dissolution of polysaccharides and lowers extraction efficiency. Thus, we measured antioxidant activity as DPPH radical scavenging assay system on six kinds of pollens inoculated with fungi to confirm the husk removal effect. The antioxidant activity was evaluated by measuring the free radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH). Briefly, extracts prepared from different extraction conditions were mixed with freshly prepared DPPH solution. After shaking, the reaction mixtures were stand for 30 min at room temperature in dark places. The radical scavenging activity was determined by measuring the absorbance at 517 nm. The relative radical scavenging activity (%) was calculated as [1-absorbance of solution with sample and DPPH / absorbance of solution with DPPH] x 100. The scavenger activities of Quercus acutissima, Actinidia arguta, Amorpha fruticosa, Oryza sativa, Chrysanthemum morifolium and Salix koreensis were 42.73±1.25, 8.98±1.46, 4.38±3.53, 36.09±0.74, 17.28±1.95 and 62.76±2.07 at 500㎕/㎖, respectively.

Bee venom is a complex mixture of toxic components that induces immediate local inflammatory and allergic responses. However, the presence and role of superoxide dismutase (SOD) in bee venom have not been previously investigated. Here, we provide the first demonstration that bee venom contains Cu,Zn SOD (SOD3), a novel extracellular component that promotes local inflammation. Bee venom SOD3 was shown to be an oxidant, rather than an antioxidant, that induces the inflammation-signaling molecule H2O2 in vivo. H2O2 plays a pathological role by triggering an immediate local inflammatory response. Furthermore, bee venom SOD3 significantly induced the activation of proinflammatory mediators (TNF-α and COX-2) and cytokines (IL-1β and IL-6) via the overproduction of H2O2 in mice. Our data demonstrate that bee venom SOD3 induced H2O2, which drives an immediate local inflammatory response, indicating a novel mechanism underlying bee venom-induced local inflammation.

Two different types of acetylcholinesterae (AChE1 and AChE2) are present in majority of insects, including the Western honey bee. Out of the two honey bee AChEs (AmAChEs), the soluble AmAChE1 with little catalytic activity is widely distributed in both neuronal and non-neuronal tissues, including fat body. In this study, to identify stresss factors that can induce AmAChE expression, we tested various conditions that honey bees can encounter in natural setting, including heat shock, cold shock, bacterial challenge (Escherichia coli and Staphylococcus aureus) and Varroa mite infestations, and evaluated their effects on AmAChE expression. Among the stress factors tested, only heat shock condition induced AmAChE expression in a dose dependet manner. This finding suggests that one function of AmAChE1 is related with thermoregulations, especially against heat shock stress in honey bees.

Honey bee swarming is a natural phenomenon that occurs when the colony encounters changes in the in-hive (i.e. population size and queen condition) and environmental conditions. To better understand the molecular basis of swarming, we conducted the transcriptomic profiles of worker bees between before swarming [pre-swarming colony (PSC)] and after swarming [swarming group (SG) and remaining group (RG)]. Based on the gene set enrichment analysis (GSEA), we predicted the biological processes associated with swarming. In addition, we analyzed the composition of cuticular hydrocarbons (CHCs) by gas chromatography-mass spectrometry and compared their profiles between different bee groups. GSEA results showed that there were a little differences between PSC and RG while many of the pathways related with metabolism and protein processing were down regulated in SG relative to PSC and RG. CHCs profiling revealed a similar CHCs composition between PSC and RG but some differences in CHCs composition (i.e. heneicosane, octacosane, octacosanol) were detected between SG and RG. These differences in gene pathway and CHC composition were discussed with respect to physiological changes and social communication.

Bee pollen have been used for centuries as a kind of traditional medicine and food diets additive due to their nutritional and biological properties. The present study was to evaluate the biological features of ethanol extracts of 18 different bee pollen (EEBP) samples collected from 16 locations of South Korea. Our results indicated most of the EEBP samples showed 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, inhibitory activities against human β-amyloid precursor cleavage enzyme (BACE-1), and antiproliferative activities towards five cancer cell lines. However, these EEBP samples did not present human acetylcholinesterase (AChE) inhibitory activities, and intestinal bacteria growth inhibitory activities.

Colony collapse disorder (CCD), a phenomenon of honeybees disappearance, has been reported since 2006. Chronic exposure to neonicotinoid insecticides, particularly imidacloprid, has been suggested to impair forager’s ability for foraging and be a main cause of CCD. Recently, it has been reported that imidacloprid induces insulin resistance in animal cell line by blocking glucose uptake. Similarly to human insulin, insulin-like peptide (ILP) of insects is involved in maintaining blood glucose contents in hemolymph by regulating the concentration of trehalose and glycogen. Therefore, we have hypothesized that sublethal concentration of neonicotinoid may affect the metabolic pathway of honey bees as well. We investigated the transcription levels of the genes involved in the insulin/insulin-like signaling (IIS) pathway, such as AmILP and AmInR, following an acute or a chronic dietary exposure of sublethal concentrations of imidacloprid to foragers. In both experiments, honeybees showed increased expression levels of ILP and InR in a dose-dependent manner. Our results suggest that sublethal dose of imidacloprid likely upregulates IIS pathway, thereby rendering honey bees to become resistant to insulin.