기후변화, 외래병해충발생, 밀원수 감소 등 여러 요인으로 인해 우리 나라의 양봉농가는 벌꿀생산량 및 농가소득이 감소하는 추세이다. 또한 밀원이 없는 시기에 가축용 사료가 아닌 고가의 식용설탕을 꿀벌의 인공사료로 급여하고 있어 농가는 ‘생산비 상승’과 ‘밀원수 부족’을 국내양봉업이 직면한 가장 큰 문제점으로 지적하고 있다. 이에 따른 양봉농가의 생산비를 절감하고 꿀벌의 생리·생태의 개선효과를 가지는 저가형 당액사료개발을 위해 농산부산물에서 추출한 천연당을 봉군에 적용시험을 추진하였다. 양파(OS-1, OS-2), 양배추(CS-1, CS-2), 바나나(Banana)부산물에서 추출한 천연당의 봉군당 평균먹이소모량(mL)은 설탕당액(대조구) 처리구와 비교시 바나나추출당액 급여구에서 5500.00±0.00, 5133.33±404.15로 유사하였다. 그러나 꿀벌의 봉세발달은 대조구에 비해 21006.7±2137, 17403.3±2257마리로 조사되어 낮은 경향을 나타내었고 수명조사 결과는 큰 차이가 없었다. 농산부산물을 이용한 천연당은 우선 꿀벌의 기호도 향상을 우선으로 개선시켜야 할 것으로 보인다.

농촌진흥청 개발 품종인 토종벌 신품종(질병저항성 우수계통)에 대해 전남지역에서의 형질특성(봉세발달, 마리당 수밀력, 청소력 등)을 조사하였다. 질병저항성 계통(RS)으로 육성된 토종벌의 유밀기 마리당 수밀력은 들어오는 일벌의 무게가 80.89±8.95mg, 나가는 일벌의 무게가 63.56±8.90mg으로 조사되었고, 비저항성계통(NRS)은 각각 83.22±1.39, 66.67±1.20mg으로 조사되어 RS에서 그 무게차가 0.7mg 더 높았으나 통계적 유의차는 없었다. Pin killed test에 의한 사충제거능력(청소력)은 제거하지 못한 유충잔존율이 RS에서 12시간과 24시간 경과시 14.00±10.39와 7.00±3.46으로, NRS에서는 20.33±14.29, 13.33±10.41%로 조사되었으며, 특히 RS와 NRS의 사충 비제거율이 24시간 경과하였을 때 4.67±2.08, 8.33±7.77%로 RS가 우수한 특성을 보였다. 신규여왕벌 입식 후 봉군세력발달은 NRS에 비하여 RS의 일벌수, 번데기 및 유충수의 발달율이 안정적이고 다소 빠르게 증가하였다.

Royal jelly (RJ) is a well-known functional and medicinal food for human health promotion. Major royal jelly proteins (MRJPs), which are the major protein components in RJ, exhibit antimicrobial activities. However, the identities of the MRJPs of RJ responsible for its antioxidant effects have remained unclear. Here, we report that honeybee (Apis cerana) MRJP 2 (AcMRJP2) acts as an antimicrobial and antioxidant agent in RJ. Using recombinant AcMRJP2, which was produced in baculovirus-infected insect cells, we established the antimicrobial and antioxidant roles of MRJP 2. AcMRJP2 bound to the surfaces of bacteria, fungi, and yeast, which then induced structural damage in the microbial cell walls and led to a broad spectrum of antimicrobial activities. AcMRJP2 protected mammalian and insect cells via the direct shielding of the cell against oxidative stress, which led to reduced levels of caspase-3 activity and oxidative stress-induced cell apoptosis, followed by increased cell viability. Moreover, AcMRJP2 exhibited DNA protection activity against reactive oxygen species (ROS). Our data indicate that AcMRJP2 could play a crucial role as an antimicrobial agent and antioxidant in RJ, suggesting that MRJP 2 is a component responsible for the antimicrobial and antioxidant activities of RJ.

Bee venom, which serves as a weapon to defend the colony from predator attacks, induces an immediate local inflammatory response that causes acute redness and swelling at the site of the sting. This venom-induced inflammation is a rapid anti-predatory defense strategy of the bee against vertebrate predators. Although acute inflammation by venom from venomous arthropods, including bees, is a typical response, how venom acutely elicits inflammatory responses remains unknown. Here, we identify a novel mechanism underlying acute inflammation and provide a rationale for the presence of superoxide dismutase (SOD3) in bee venom. In mouse models, paradoxically, SOD3 in bee venom (bvSOD3) acts as a reactive oxygen species (ROS)-based harm-inducing system to promote acute inflammation. Exogenous bvSOD3 rapidly induced overproduction of H2O2 through endogenously produced superoxide by venom components, such as melittin and phospholipase A2 (PLA2), which then upregulated the expression of proinflammatory genes and promoted the acute inflammatory response. Furthermore, a more severe noxious effect by bvSOD3 elevated a type 2 immune response, and bvSOD3 immunization protected against bvSOD3-mediated toxicity. Our findings that bvSOD3 promotes an acute inflammatory response and induces a protective immune response against inflammation may offer a new approach in venom therapy/immunotherapy.

Honeybee (Apis mellifera) egg-yolk protein vitellogenin (Vg) plays roles in immunity, antioxidation, and life span beyond reproduction, but it also acts as an allergen Api m 12 in venom. Here we established antimicrobial and antioxidant roles of honeybee Vg in the body and venom. Using the cDNA encoding Vg identified from Asiatic honeybee (A. cerana) workers, recombinant A. cerana Vg (AcVg) protein of approximately 180 kDa was produced in baculovirus-infected insect cells. In A. cerana worker bees, AcVg was expressed in the fat body and venom gland and was present in the secreted venom. AcVg induced structural damage in microbial cell walls via binding to microbial surfaces and exhibited antimicrobial activity against bacteria and fungi. AcVg protected mammalian and insect cells against oxidative damage through direct shielding of cell membranes. Interestingly, AcVg exhibited DNA protection activity against reactive oxygen species (ROS). Furthermore, the transcript level of AcVg was upregulated in the fat body, but not in the venom gland, of worker bees with antimicrobial peptides and antioxidant enzymes in response to microbial infection and oxidative stress. Our data indicate that AcVg is involved in innate immunity upon infection and in a defense system against ROS, supporting a crucial role of honeybee Vg as an antimicrobial and antioxidant agent in the body and venom.

The Acoptolabrus changeonleei Ishikawa et Kim, 1983 (Coleoptera: Carabidae), has been listed as an endangered insect in South Korea. The complete mitochondrial genome of the species was 16,831 bp with a typical set of genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and one non-coding region, with the arrangement identical to that observed in most insect genomes. Phylogenetic analyses with concatenated sequences of the 13 PCGs and 2 rRNA genes, using the Bayesian inference (BI) and maximum-likelihood (ML) methods, placed A. changeonleei as a sister to the within-subfamilial species Damaster mirabilissimus in Carabinae, with the highest nodal support by both analyses.

Bee venom contains a variety of peptide constituents, including low-molecular-weight protease inhibitors. While the putativelow-molecular-weight serine protease inhibitor Api m 6 containing a trypsin inhibitor-like cysteine-rich domain was identifiedfrom honeybee (Apis mellifera) venom, no anti-fibrinolytic or anti-microbial roles for this inhibitor have been elucidated.In this study, we identified an Asiatic honeybee (A. cerana) venom serine protease inhibitor (AcVSPI) that was shownto act as a microbial serine protease inhibitor and plasmin inhibitor. AcVSPI was found to consist of a trypsin inhibitor-likedomain that displays ten cysteine residues. Interestingly, the AcVSPI peptide sequence exhibited high similarity to the putativelow-molecular-weight serine protease inhibitor Api m 6, which suggests that AcVSPI is an allergen Api m 6-like peptide.Recombinant AcVSPI was expressed in baculovirus-infected insect cells, and it demonstrated inhibitory activity against trypsin,but not chymotrypsin. Additionally, AcVSPI has inhibitory effects against plasmin and microbial serine proteases; however,it does not have any detectable inhibitory effects on thrombin or elastase. Consistent with these inhibitory effects, AcVSPIinhibited the plasmin-mediated degradation of fibrin to fibrin degradation products. AcVSPI also bound to bacterial andfungal surfaces and exhibited anti-microbial activity against fungi as well as gram-positive and gram-negative bacteria. Thesefindings demonstrate the anti-fibrinolytic and anti-microbial roles of AcVSPI as a serine protease inhibitor.

Major royal jelly proteins (MRJPs) are important protein components of bee royal jelly (RJ) and exhibit various biologicaland pharmacological activities. The antimicrobial activities of royalisins and the jelleines contained within MRJP 1 andMRJP 2 in RJ have been elucidated. However, the antimicrobial effects of other bee RJ MRJPs remain largely unknown.In this study, we demonstrated that the Asiatic honeybee (Apis cerana) MRJP 4 (AcMRJP4) exhibits antimicrobial activitiesagainst bacteria, fungi, and yeast. Recombinant AcMRJP4 was expressed as a 63-kDa protein in baculovirus-infected insectcells. However, some of the recombinant AcMRJP4 proteins were cleaved into two fragments (i.e., 48-kDa (AcMRJP4-48)and 15-kDa (AcMRJP4-15) proteins) by the proteolytic cleavage of the C-terminus of the recombinant AcMRJP4. Interestingly,AcMRJP4, AcMRJP4-48, and AcMRJP4-15 exhibited antimicrobial activities, with AcMRJP4-15 exhibiting the highestantimicrobial activity, followed by AcMRJP4. AcMRJP4-15, which is a hydrophilic peptide with 88 amino acid residuesthat contains a high content of Asn and positively charged amino acids, induced structural damage in the cell walls ofthe assayed bacteria, fungi, and yeast. Altogether, our data demonstrated that AcMRJP4 functions as an antimicrobial agent.

We performed temperature dependent current-voltage (I-V) measurements to characterize the electrical properties of Au/Al2O3/n-Ge metal-insulator-semiconductor (MIS) diodes prepared with and without H2O prepulse treatment by atomic layer deposition (ALD). By considering the thickness of the Al2O3 interlayer, the barrier height for the treated sample was found to be 0.61 eV, similar to those of Au/n-Ge Schottky diodes. The thermionic emission (TE) model with barrier inhomogeneity explained the final state of the treated sample well. Compared to the untreated sample, the treated sample was found to have improved diode characteristics for both forward and reverse bias conditions. These results were associated with the reduction of charge trapping and interface states near the Ge/Al2O3 interface.

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.

Detergency and surface active properties of mixed anionic surfactants with amphoteric and nonionic were investigated. Sodium dodecyl sulfate (SDS) and ammonium dodecyl sulfate (ADS) as anionic surfactants and cocamidopropyl betaine (CAPB) as an amphoteric surfactant were used. Nonionic surfactants, which are butyl glucoside (BG), octyl glucoside (OG), decyl glucoside (DG), lauryl dimethylamine oxide (AO) and saponin were also used. To study the synergy effects of mixed SDS/ADS anionic surfactant systems, amphoteric and nonionic surfactants were added into the mixed anionic surfactants. Investigated properties of surfactant mixtures were critical micelle concentration (CMC), surface tension (γ), wettability. In addition, based on these properties, detergency of each sample was examined. Surfactant mixtures are anionics (SDS/ADS), anionic/amphoteric/nonionic (SDS/ADS/CAPB/ saponin), and anionic/nonionic (SDS/ADS/BG/saponin, SDS/ADS/OG/saponin, SDS/ADS/ DG/saponin, and SDS/ADS/AO/saponin). With the addition of amphoteric and nonionic to mixed anionic surfactants, CMC and γ were decreased. Addition of CAPB, which is amphoteric, showed the best property at CMC and γ. Furthermore, as the chain length of hydrocarbon in alkyl glucosides was increased, the CMC and γ were enhanced. However, the wettability did not exactly match up with CMC and γ. The surfactant mixture, which contained DG, showed the best performance at wetting time. Detergency was measured at various temperatures (15 oC, 30 oC, 50 oC). The cleaning performance was enhanced by increasing washing temperature. Moreover, detergency was influenced by not only CMC and γ but also wettability. Although CMC and γ were not minimum at surfactant mixture that included DG, the best cleaning performance showed in that sample.

Bee venom contains a variety of peptide constituents that have various biological, toxicological, and pharmacological actions. However, the biological actions of secapin, a venom peptide in bee venom, remain largely unknown. Here, we provide the first evidence that the Asiatic honeybee (Apis cerana) secapin (AcSecapin-1) exhibits anti-fibrinolytic, anti-elastolytic, and anti-microbial activities. AcSecapin-1 functions as a serine protease inhibitor-like peptide that has inhibitory effects against plasmin, elastases, microbial serine proteases, trypsin, and chymotrypsin. Consistent with these functions, AcSecapin-1 inhibited the plasmin-mediated degradation of fibrin to fibrin degradation products, thus indicating the role of AcSecapin-1 as a clotting factor. AcSecapin-1 also inhibited both human neutrophil and porcine pancreatic elastases. Furthermore, AcSecapin-1 exhibited anti-microbial activity against fungi and Gram-positive and Gram-negative bacteria. Taken together, our data demonstrated that AcSecapin-1 has a multifunctional role as an anti-fibrinolytic agent, an anti-elastolytic agent, and an anti-microbial peptide, and our data suggested novel functions for the biological actions of the bee venom peptide, secapin.

Inhibitor cysteine knot (ICK) peptides exhibit ion channel blocking, insecticidal, and antimicrobial activities, but currently, no functional roles for bee-derived ICK peptides have been identified. In this study, a bee (Apis cerana) ICK peptide (AcICK) that acts as an antifungal peptide and as an insecticidal venom toxin was identified. AcICK contains an ICK fold that is expressed in the epidermis, fat body, or venom gland and is present as a 6.6-kDa peptide in bee venom. Recombinant AcICK peptide (expressed in baculovirus-infected insect cells) bound directly to Beauveria bassiana and Fusarium graminearum, but not to Escherichia coli or Bacillus thuringiensis. Consistent with these findings, AcICK showed antifungal activity, indicating that AcICK acts as an antifungal peptide. Furthermore, AcICK expression is induced in the fat body and epidermis after injection with B. bassiana. These results provide insight into the role of AcICK during the innate immune response following fungal infection. Additionally, we show that AcICK has insecticidal activity. Our results demonstrate a functional role for AcICK in bees: AcICK acts as an antifungal peptide in innate immune reactions in the body and as an insecticidal toxin in venom. The finding that the AcICK peptide functions with different mechanisms of action in the body and in venom highlights the two-pronged strategy that is possible with the bee ICK peptide.

We have used bulked segregant analysis to screen the strain-specific DNA marker associated thermophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 550 bp was amplified with OP-S07 primer from the thermophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-S07-1. The PCR analysis with the OP-S07-1 primer showed that this SCAR marker clearly distinguish the thermophilic strains from the control strains.

Insect-derived Kazal-type serine protease inhibitors exhibit thrombin, elastase, plasmin, proteinase K, or subtilisin A inhibition activity, but so far, no functional roles for bee-derived Kazal-type serine protease inhibitors have been identified. In this study, a bee (Apis cerana) venom Kazal-type serine protease inhibitor (AcKTSPI) that acts as a microbial serine protease inhibitor was identified. AcKTSPI contained a single Kazal domain that displayed six conserved cysteine residues and a P1 threonine residue. AcKTSPI was expressed in the venom gland and was present as a 10-kDa peptide in bee venom. Recombinant AcKTSPI Kazal domain (AcKTSPI-Kd) expressed in baculovirus-infected insect cells demonstrated inhibitory activity against subtilisin A (Ki 67.03 nM) and proteinase K (Ki 91.53 nM), but not against α-chymotrypsin or typsin, which implies a role for AcKTSPI as a microbial serine protease inhibitor. However, AcKTSPI-Kd exhibited no detectable inhibitory effects on factor Xa, thrombin, tissue plasminogen activator, or elastase. Additionally, AcKTSPI-Kd bound directly to Bacillus subtilis, B. thuringiensis, Beauveria bassiana, and Fusarium graminearum but not to Escherichia coli. Consistent with these findings, AcKTSPI-Kd showed antibacterial activity against Gram-positive bacteria and antifungal activity against both plant-pathogenic and entomopathogenic fungi. These findings constitute molecular evidence that AcKTSPI acts as an inhibitor of microbial serine proteases. This paper provides a novel view of the antimicrobial functions of a bee venom Kazal-type serine protease inhibitor.

The pear psylla, Cacopsylla pyricola Foerster (Homoptera: Psyllidae), is a serious insect pest of commercial pear crops. The species, which resides on pear trees throughout its life cycle, is rapidly spreading in some regions of the world. Given the life cycle, it is unclear how such a rapid spread has been facilitated. Presently, the population genetic structure of the species including genetic diversity and gene flow was studied to understand the nature of dispersal and field ecology of the species. Pear psylla was collected from several pear orchards in Korea. The 658-bp region of mitochondrial COI gene and the 716-bp long complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA were sequenced. Unlikely other previously studied insect pests, the COI-based genetic diversity of the pear psylla was extremely low (maximum sequence divergence of 0.15%). This finding allowed us to conclude that the species may have been introduced in Korea relatively recently, possibly with the phenomenon of genetic bottlenecks. ITS2 sequence-based analyses of phylogeny, population differentiation, gene flow, and hierarchical population structure all concordantly suggested that the pear psylla populations in Korea are neither genetically isolated nor hampered for gene flow. These genetic data are concordant with the dispersal of an overwintering winterform morph outside the non-pear habitat in the fall and the possibility of subsequently longer distant dispersal.

To control pest of Brassicaceae leafy vegetable(leaf broccoli, Red Mustard Leaf, Tatsoi) which grows at vinyl house in IKSAN, Jeollabuk-do, the non-treatment was set as negative control, and treatment was divided into Parasitic natural treatment group and general treatment group. And incidence density of pest was surveyed and control effect was analyzed. As a result, in case of spring plants, the leaf damage ratio was decreased by 31% in Leaf broccoli, 30% in Red mustard leaf and 27% in Tatsoi compared to untreatment. In case of autumn plants, it was decreased by 32%, 41% and 17% respectively. The key pests were Plutella xylostella, Pieris rapae, Myus persicae Sulzer, Thrips palmi and Striped cabbage flea-beetle. Compared with the untreatment, the incidence density of each was significantly controlled. Other pests include Spodoptera exigua, Macdunnoughia purissima, Macdunnoughia purissima which showed high incidental density sometimes. In case of spring plants, the number by treatment was increased by 117% in Leaf broccoli, 85% in Red mustard leaf and 1,000% in Tatsoi. In autumn plants, it was increased by 132%, 257% and 1,077% respectively. The used Parasitic natural and eco-friendly materials were Cotesia glomerata, lacewing, ladyburg, Orius laevigatus, Encarsia formosa, Entomopathogenic nematode, Chungjigi and Togkaki. During early development of each pest, the Parasitic naturals were grazed 2~3 times at the interval of 7~10days. During the peak time, eco-friendly materials were sprayed 1~2 times. Based on this, comprehensive management model was drafted by period for each Brassicaceae vegetables pest.

To control Thrips tabaci in Korean leek and green onion which grow at vinyl house in IKSAN, Jeollabuk-do, the non-treatment was set as negative control. In general treatment group, 120 mesh gauze was installed on the side window after planting(May, 2nd) and, in spring, Orius strigicollis (1.0/m2) was grazed 3 times every 7 days from mid of May which is early development stage and In the mid of June, which is the peak stage, plant extracts were sprayed one time. In autumn, O. strigicollis was grazed 3 times every 7 days from mid of September which is early development stage, and in the mid of October, which is the peak stage, plant extracts were sprayed one time. The result shows that the leaf damage ratio was decreased by 22% in Korean leek and by 27% in green onion compared to the untreatment. And the control value of Thrips tabaci shows 78.7% in korean leak and 90.6% in green onion. The density control effect of Thrips tabaci was significantly controlled under max 6.2/plant in general treatment group compared max 25/plant in the untreatment and this result was similar in green onion. The yields by general treatment was increased by 85% in Korean leek and 56% in green onion, compared with non-treatment yields, which was 900kg/10a in Korean leek and 1,287kg/10a in green onion.