메뚜기목 메뚜기과에 속하는 풀무치(Locusta migratoria)는 강한 집단행동 습성과 다양한 먹이식물에 대한 높은 유연성을 가진 대형 메뚜기이다. 이런 풀무치를 실내에서 사육하기에 가장 적합한 온도를 조사하였다. 실험온도는 25℃, 27℃, 30℃, 33℃ 등 4개구로 설정하였고, 12L/12D, 65% RH, 1,000 lux 이상의 사육조건하에서 밀을 먹이로 실험하였다. 풀무치의 영별 발육특성을 조사한 결과, 1령-5령 약충까지의 발육기간은 30℃가 26.4±2.0일로 25℃ (46.9±1.9일)보다 약 1.8배, 27℃(31.8±2.5일)와 33℃(30.7±2.6일)보다는 약 1.2배 단축되었다. 약충의 체중, 두폭, 체폭, 체장 등 발육 특성은 30℃가 가장 좋았고 그 다음은 33℃, 27℃, 25℃순이었다. 성충의 교미는 주로 09-10시와 13-14시에 가장 많이 이루어졌다. 우화-교미까지의 일수는 30℃가 10.2±1.5일로 25℃(23.8±13.7일), 27℃(19.3±10.2일), 33℃(12.0±7.3일)보다 약 1.8일-13.6일 빨랐다. 포토 당 산란된 난괴의 수는 평균 1.1개로 나타났다. 이상의 결과들로 볼 때 풀무치를 사육하기에 가장 적합한 온도는 30℃인 것으로 판단된다.

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.

생대추가 신소득 작목으로 부각되면서 재배지가 확대되고 있으나 우리나라 장마기와 최근 이상기후로 인해 대추 수분·수정 및 착과에 문제 가 발생하고 있는 실정이다. 이에 생대추 재배를 위해 도입된 비가림하우스에서 서양종 꿀벌과 서양뒤영벌을 이용하여 화분매개곤충의 활동 특성 과 착과 효율에 어떠한 영향을 미치는지 구명하고자 본 시험을 수행하였다. 대추 복조 품종을 대상으로 비가림하우스에 망실을 씌워 서양종 꿀벌 과 서양뒤영벌을 2016년 6월 1일부터 7월 25일까지 약 55일정도 방사하였다. 서양종 꿀벌은 주로 오후에 활동이 많았으나 서양뒤영벌의 활동은 시간에 관계없이 다소 균일하였다. 대추 신초 가지의 착과율을 조사한 결과 방화곤충을 차단한 무처리 5.5% 대비 서양종 꿀벌은 10.2%, 서양뒤 영벌은 8.9%로 대추 비가림하우스내 화분매개곤충 활용시 착과량 증진으로 농가 소득증대에 기여할 것으로 생각된다.

We investigated pollination properties of bumblebees in a high temperature condition i. e. a greenhouse where onions(Aliumcepa) were cultivated. Foraging activity of bumblebee workers decreased at temperature over 33 ℃ and was negativelycorrelated with colony temperature. Fanning behavior was observed at temperature over 30 ℃ and was positively correlatedwith colony temperature. There was negative correlation between the number of bees fanning and bees foraging. We expectedthat foraging activity would be affected by colony temperature and fanning behavior. The temperature of the coloniesthat were ventilated using the cooler fan was 3 ℃ lower than the control colonies, and the rate of fanning behaviorwere 9 times lower in ventilated colonies than in the control colonies. Foraging activity and survival rates of workerbees in ventilated colonies doubled compared to the control colonies

We have surveyed the current status of insect pollinator use for horticultural crops in 2016. The use rate and farmnumber of insect pollinators for 26 horticultural crops were 25.8% and 55,208, respectively. The colony number of insectpollinators used in this survey was 479,777, which include 344,690 for honeybees, 119,104 for bumblebees, 2,415 formason bees, 1,317 for flies, and 2,415 for the combination of bumblebees, honeybees, and mason bees. The use rateof insect pollinators was 59.4% for 11 vegetable crops and the colony number of insect pollinators used for 11 vegetablecrops was 449,287. The colony number of insect pollinators used for 15 fruit tree crops was 30,290, which include honeybees(66.3%), bumblebees (20.2%), mason bees (8.0%), flies (1.6%), and the combination (3.9%) of bumblebees, honeybees,and mason bees. Together, farms of 98% showed positive effect for the use of insect pollinators and most of farms (97.0%)planed for the continuous use of insect pollinators

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 collected four species of Osmia spp. consisting of O. cornifrons, O. pedicornis, O. taurus and O. satoi in Yeongwol, Bonghwa and Yechon locations. O. pedicornis was the dominant species, which accounts for 59.6% to 76.4% in the three locations. The sex ratio of Osmia spp. were from 1:1 to 1: 2.8 in O. cornifrons and from 1:2.8 to 1: 5.5 in O. pedicornis. The sex ratio of O. cornifrons and O. pedicornis was different in 3 locations. The female and male cocoons of O. cornifrons were the long elliptical-shaped and grey-brown color. The female and male cocoons of O. pedicornis were the long elliptical-shaped and polished dark-brown color. In both species of O. cornifrons and O. pedicornis, the cocoon size of the females was significantly larger than that of the male. Between those two species, the latter had a relatively bigger size than the former.

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.

In the age of sexual maturity of queen, mating occurred at 6.7% immediately after eclosion and it was the highest as 85.0% at 10 days after emergence. However, a remarkable decrease was occurred at 20 days after the emergence. With regard to oviposition rate, the highest rate was observed as 81.3-81.8% at 6 to 10 days of eclosion. The rate of colony foundation and progeny-queen production were the highest as 43.8% and 37.5%, respectively, at 8 days of eclosion. In the age of sexual maturity of male, mating rate was as high as 38.3% immediately after the eclosion, the highest as 80.0% at 25 days of eclosion, and 76.6% at 8 days of eclosion. The oviposition rate was the highest as 76.9% at 6 days of eclosion, and decreased to 75.0% at 8 days and 72.7% at 10 days of eclosion. Summarized, our results indicate that sexual maturity for mating of B. terrestris is most favorable 6-8 days after eclosion for queen and 6-10 days after for male.

The purpose of this study was to identify the new fungal disease of Scopendra subspinipes mutilans (Chilopoda: Scolopendridae) from Jeju island in Korea. Scopendra subspinipes mutilans are also known the Chinese red head and average 20cm in length. It is used for traditional medicine for skin-disease and tumor and S. s. mutilans is recently raised in Jeju island to use healing properties. We isolated fungi from surface of infected S. s. mutilans and incubated in SDBA at 24℃ for 14days, and then identified the morphology of the fungi by light and electron microscopy. For the specific diagnosis of Metarhizium spp. in S. s. mutilans, 18srRNA of fungi was amplified by primers ITS4_MF and LR3_MR and sequenced by using the BLAST server at the National Center for Biotechnology Information. The result of the 18srRNA sequencing alignment was Metarhizium anisopliae (99.9%). The entomopathogenic fungi, M. anisopliae would be fatal cause of higher mortality to S. s. mutilans, and it is needed in vivo assay to confirm the pathogenic activities of M. anisopliae to S. s. mutilans.

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.

We investigated pollination and foraging activities of honeybee (Apis mellifera L.) and bumblebee (Bombus terrestris L.) during flowering season of the asian pear (Pyrus pyrifolia N.) under different weather conditions. There was no significant statistical difference about the pollination activities of two species. However, the pollination activities of bumblebee were more active than those of honeybee under low temperature and rainfall period. The activities of honeybee and bumblebee were more influenced by temperature than other factors (i.e. illumination and wind velocity). Honeybee was more sensitive to temperature and illumination than bumblebee. At low temperatures (<20℃) on cloudy days (<30,000 lux) with a certain wind velocity (>4.0 m/s), the pollination activity of the honeybee was lower twice than that of bumblebee.

Distillers dried grain (DDG) and makgeolli spent grain (MSG) are agricultural by-product to produce alcoholic beverage. However, they are known to contain enough nutrients. Mealworm is a promising insect resource for an animal feed ingredient as well as alternative human food. With low cost, DDG and MSG were investigated as a feed ingredient for rearing high quality mealworms. DDG and MSG were mixed with wheat bran and compared to control feed (only wheat bran) for its effects on larval survivorship, larval weight, duration for larval development, pupation rate, and pupal weight. Adding DDG on wheat bran showed positive results for larval weight, duration for larval developmental period, and pupation rate. However, adding MSG made longer duration for larval development, but it also improved larval weight, pupal weight with more than 90% pupation rate. We confirmed that adding 30～50% of DDG or MSG to conventional wheat bran have a strong potential to replace the conventional wheat bran insect feed for quality insect production.

Serine protease inhibitors play a critical role in physiological processes and immune responses by regulating serine protease activities. Here we describe the molecular cloning and antimicrobial activities of a serine protease inhibitor from the mason bee, Osmia cornifrons (OcSPI). OcSPI consists of 405 amino acid residues and contains a potential reactive center loop (RCL) region in its C-terminus. Recombinant OcSPI was produced as a 64-kDa glycoprotein in baculovirus-infected insect cells and exhibited inhibitory activity against chymotrypsin. Additionally, OcSPI demonstrated inhibitory activity against microbial serine proteases, such as subtilisin A and proteinase K, but not against tissue plasminogen activator, thrombin, or plasmin. Recombinant OcSPI bound directly to Escherichia coli, Bacillus subtilis, and Beauveria bassiana and exhibited antimicrobial activity against both bacteria and fungi. Our results demonstrated the antimicrobial functions of OcSPI and suggest a role for OcSPI in the immune response of O. cornifrons.

Osmia cornifrons is a cavity-nesting solitary species used as an apple pollinator in Korea. To elucidate the developmental characteristics of O. cornifrons, we investigated its development from the egg to adulthood, including a dormant prepupal phase and mating through indoor rearing (25 °C, 65% R.H.). The egg durations of the female and male bees were 3.6 ± 0.8 days and 3.1 ± 1.3 days, respectively. During larval development, the head widths of the 1st to 5th instars ranged from 0.7 ± 0.1 mm to 1.3 ± 0.1 mm. The peak of the growth in head width was the 2nd instar. The larval lengths ranged from 3.7 ± 0.6 mm to 13.6 ± 1.3 mm. The peak of growth was the 4th instar. The larval weights ranged from 4.5 ± 1.2 mg to 78.3 ± 16.1 mg. The peak of growth was the 3rd instars. The total larval durations of from the 1st to 5th instars for the females and males were 14.0 ± 6.0 days and 13.2 ± 5.8 days, respectively. The spinning durations of the females and males were 2.2 ± 0.7 days and 2.3 ± 0.8 days, the prepupation durations were 55.5 ± 5.9 days and 55.8 ± 2.9 days, and the pupation durations were 26.4 ± 2.1 days and 25.3 ± 2.3 days, respectively. The average longevity of the female adults and male adults was 21.8 ± 8.7 days and 24.4 ± 12.4 days, respectively. The total duration of from the egg to an adult bee of the O. cornifrons females and males was 123.5 days and 124.1 days, respectively. Mating consisted of the three following phases: the precopulatory (courtship and attempting copulation), copulation and postcopulatory phases. The mating times of the precopulatory, copulation and postcopulatory phases were 159.6 ± 288.9, 8.4 ± 7.1, 12.9 ± 4.5, and 198.8 ± 69.8 seconds.

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.

The honeybee inhibitor cysteine knot (ICK) peptide acts as an antifungal peptide and insecticidal venom toxin. However, the ICK peptide from bumblebees has not been characterized. Here, we report the molecular cloning and antifungal activity of a bumblebee (Bombus ignitus) ICK peptide (BiICK). We identified a BiICK that contains an ICK fold. The BiICK was expressed in the epidermis, fat body, and venom gland of B. ignitus worker bees. A 6.7-kDa recombinant BiICK peptide was expressed in baculovirus-infected insect cells. Recombinant BiICK peptides directly bound to Beauveria bassiana, Ascosphaera apis, and Fusarium graminearum, but they did not bind to Escherichia coli, Paenibacillus larvae, or Bacillus thuringiensis. Consistent with this finding, BiICK exhibited antifungal activity against fungi. These results demonstrate that BiICK acts as an antifungal peptide.

The Osmia cornifrons plays an important role in pollinating fruit trees, such as apple trees. To better understand diapause and oviposition in O. cornifrons, we investigated the correlation between the ovarian development and secretion level of OcVg protein in hemolymph. During ovarian development in wintering the number of oocytes progressively increased in comparison with the length of the ovaries and the oocytes. After emergence, the oocyte and ovary sizes developed until 6 days after emergence and declined after 6 days, but the number of oocytes decreased gradually. The secretion level of OcVg protein in hemolymph revealed that during wintering, the secretion level increased from 1 month to 2 months and then stagnated after 2 months. After diapause, the secretion level increased gradually until day 6 of the newly emerged adult from cocoon stage, and thereafter gradually declined, remaining detectable until day 30 of the adult stage. The correction analysis between ovarian development and OcVg secretion level in hemolymph found that in wintering, the number of oocytes was positively correlated to OcVg secretion level. After diapause, the ovary and first oocyte lengths and the number of oocytes showed significant changes in OcVg secretion level and positively correlated with OcVg secretion level, respectively. These results suggest that there is a significant interaction between ovarian development and the secretion level of OcVg protein and the pattern of ovarian development and secretion of OcVg protein are stage-specific in the O. cornifrons female.

Bees of the genus Osmia are cavity-nesting solitary species that play important roles as pollinators. Nest traps are one of the most common methods to study Osmia bees. To elucidate the optimal environmental conditions of nesting sites, we investigated the effects of location, direction, altitude, and sites of nest traps on the rate of trap-nesting Osmia spp. During the collection period, the average rate of trap-nesting Osmia spp. collected within 90 days after the installation of traps was 17.0±20.0%. This percentage was 2.7-fold higher than that of trap-nesting bees in 30 days after the installation of traps. The Jeongseon location exhibited the highest rates of trap-nesting Osmia spp. collected in 30 and 90 days, representing 11.1 ±17.6% and 23.2±22.5%, respectively. The direction of the nest traps did not affect the rate of trap-nesting Osmia spp. The altitude ranges of the traps were 0-199 m, 200-399 m, 400-599 m, 600-799 m and over 800 m. Interestingly, the altitude range of 600-799 m showed the highest rate of trap-nesting bees, which was 40.4±3.9%. Higher altitudes seemed to correspond to higher rates of trap-nesting bees. With regards to the sites where the nests were placed, the rate of trap-nesting bees in a mud wall of an old house was 45.1±25.2%, which was 3-fold higher than that of a nest in an apple orchard. The flowering plants collected at different locations during the trap-nesting activity of Osmia bees belonged to 18 families and 34 species. In conclusion, the rates of trap nests colonized by Osmia spp. were affected by altitude, site, and plant diversity.