We used the three kinds of mosquito traps (Black-hole with UV light, CO2-baited Mos-hole with the newly developed attracting-solvent, CO2-baited DMS; Digital Mosquito Monitoring System) to know their collecting efficiency for the female mosquitoes in Korean rural areas. The Black-hole mosquito trap caught many kinds of insects including only few female mosquitoes. The Black-hole trap has the UV-light and the light seemed to attract other terrestrial and aquatic insects, such as the common flies, May flies, and the stone flies. Even though the trap was developed to collect mosquitoes, the trap caught only few of female mosquitoes less than 1% of all insects caught. Their selective efficiency to collect the female mosquitoes was relatively lower than other two kinds of traps. The Mos-hole and CO2-baited DMS traps had the collecting efficiency of over 80% to collect the female mosquitoes. The two traps caught the relatively lower number (less than 3% of total insects) of other insects, such as few Coleoptera and Diptera, and their collecting efficiency for the female mosquitoes was very higher. Generally speaking, mosquitoes disliked the UV light but they relatively preferred CO2 gas including the attracting-solvent. They had also been attracted the acidic solvent with CO2 gas. If we could use the efficient and selective mosquito traps with the fully understanding about the mosquito habits, we could assume that we can keep the biodiversity high around the mosquito habitats as well as to save money for the insect pest control.

This snag-dwelling arthropod community study was conducted for the ecological evaluation of dead woods at Korean fir stand in Mt. Woonak in Pocheon-si, Gyeonggi-do, from April 2010 to August 2011. We put a windows trap and an emergence trap on the trunk of each snag, and we selected the six snags during the study periods. We collected 3930 individuals (5 class, 21order, 52 families) but we didn’t include the number of unidentified larva. We separated those individuals into the three functional groups and we found out the proportion and number of each functional group from the total individuals: herbivores (27.6%, 1083) predators (10.9%, 430), detritivores (61.4%, 2413), etc(0.1%, 4). We found out the proportion and number of each taxon group for herbivores, Armadillidae (0.15%, 6), Acarina (2.57%, 101), Psocoptera (0.25%, 10), Hemiptera (0.46%, 18), Mecoptera (0.05%, 2), Hymenoptera (not ant) (5.14%, 202), Aphididae (3.82%, 150), Cicadellidae (0.4%, 16), Curculionidae (3.61%, 142), Chrysomelidae (0.23%, 9), Elateridae (3.36%, 132), Erotylidae (3.16%, 124), Nitidulidae (2.6%, 102), Pyrochoroidae (0.08%, 3), Scarabaeidae (0.31%, 12), and Cetoniidae (0.13%, 5). Predators were consisted of the following taxa groups: Araneae (2.9%,114), Chilopoda (0.31%, 12), Formicidae (4.25%, 167), Carabidae (0.08%, 3), Staphylinidae (1.09%, 43), Cleridae (0.05%, 2), Pselaphidae (0.1%, 4), Colydiidae (0.38%, 15), Harpalidae (0.1%, 4), Histeridae (0.36%, 14), and Dermestidae (0.8%, 31). Detritivores were consisted of the following taxa groups: Millipedes (0.92%, 36), Archaeognatha(1.6%, 63), Diptera (7.81%, 307), Collembola (35.47%, 1394), Protura (0.03%, 1), Dermaptera (0.1%, 4), Tettigoniidae (0.08%, 4), Raphiidophoridae (0.03%, 1), Ipidae (14.12%, 555), Silphidae (0.15%, 6), Cuculidae (0.15%, 6), Cerambycidae (0.38%, 15), Oedeeridae (0.03%, 1), Lucamnidae (0.03%, 1), Stenotrachelidae (0.05%, 2), Buprestidae (0.13%, 5), Tenebrionidae (0.23%, 9), and Mordellidae (0.1%, 4), etc. Conclusively, the snag plays an important roll as the diverse arthropods’ habitats in the Korean fir forest ecosystem.

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.

Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.

Classical swine fever virus (CSFV) envelope glycoprotein E2 is the main target for inducing neutralizing antibodies and protective immunity in swine. Here, we report a novel strategy forthe large-scale production of a CSFV E2 subunit vaccine that demonstrates a high immunogenic capability in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (CSFV E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed an approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Mice that were immunized with the granule form of recombinant polyhedra or the soluble form of the fusion protein elicited CSFV E2 antibodies, which indicated that the recombinant polyhedra carrying CSFV E2ΔC were immunogenic. The virus neutralization test showed that the serum from mice that were treated with recombinant polyhedra or the soluble form of the fusion protein contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen and that the recombinant polyhedra containing CSFV E2ΔC as a granule antigen can be used as a potential subunit vaccine against CSFV.

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainlyinvolved in the xenobiotic metabolism and protection against oxidative damage. Most studies of GSTs in insects have been focused on their role in detoxifying exogenous compounds in particular insecticides. Here, we show the expression profiles of GSTs of the bumblebee Bombus ignitus in response to oxidative stress. We identified a sigma-class GST from B. ignitus (BiGSTS). The BiGSTSgene consists of 4 exons that encode 201 amino acids. Comparative analysis indicates that the predicted amino acid sequence of BiGSTS shares a high identity with the sigma-class GSTs of hymenopteran insects such as Apis mellifera (70% protein sequence identity) and Solenopsis invicta (59% protein sequence identity). Tissue distribution analyses showed the presence of BiGSTS in all tissues examined, including the fat body, midgut, muscle and epidermis. The oxidative stress responses analyzed by quantitative real-time PCR showed that under H2O2 overload, BiGSTS and BiGSTD (identified in our previous study) were upregulated in all tissues examined, including the fat body and midgut of B. ignitus worker bees. Under uniform conditions of H2O2 overload, the expression profile of GSTs and other antioxidant enzyme genes, such as phospholipid-hydroperoxide glutathione peroxidase (Bi-PHGPx) and peroxiredoxins (BiPrx1 and BiTPx1), showed that other antioxidant enzyme genes are acutely induced at 3 h after H2O2 exposure, whereas BiGSTS and BiGSTD are highly induced at 9 h after H2O2 exposure in the fat body of B. ignitus worker bees. These findings indicate that GSTs and other antioxidant enzyme genes in B. ignitusare differentially expressed in response to oxidative stress. Taken together, our findings indicate that BiGSTS and BiGSTD are oxidative stress-inducible antioxidant enzymes that may play a role in oxidative stress response.

This study was estimated for cabbage aphid, Brevicoryne brassicae in Brassica campestris L. var. rapa (L.) Hartm. in order to institute of Economic injury levels(EILs). B. brassicae was innoculated on April 29, in differently 0, 5, 10, 20 and 40 adults per ten plant, respectively. After inoculated of B. brassicae, the density was increased until harvest ing gradually in all plots except non innoculate plot. and Higher inoculation density were increased higher than lower inoculation density. Percentage of damage leaf was higher in plots with higher initial aphid density than in plots with lower initial aphid density. And the leaf weight of commodity were decreased in higher initial aphid density. The decreasing rates of leaf weight of commodity was increased with increasing initial aphid density. The relationship between initial B. brassicae densities and the decreasing rates of leaf weight of commodity was well described by a linear regression, Y=0.8416X-3.5147, R2=0.94. Based on the relationship, the number of adults per 10 plant which can cause 5% loss of yield was estimated to be approximately 10.1. And EILs was estimated to be approximately 1.0 adults/plant in late April.

Peptidoglycan recognition proteins (PGRPs) are pattern recognition molecules of the innate immune system that recognize peptidoglycan, a unique cell wall component of bacteria. Here we cloned and characterized PGRP-S from the bumblebee Bombus ignitus (BiPGRP-S). The BiPGRP-S gene consists of four exons encoding 194 amino acid residues. Comparative analysis indicates that the predicted amino acid sequence of BiPGRP-S shares high identity with enzymatically active PGRP-S proteins and contains the amino acids required for amidase activity. BiPGRP-S in B. ignitus worker bees is constitutively expressed in boththe fat body and epidermis, and it is secreted into the hemolymph. Quantitative real-time PCR assays revealed that in both the fat body and epidermis, the BiPGRP-S gene is highly induced by an injection of Bacillus thuringiensis. In addition, recombinant BiPGRP-S expressed as a 19-kDa protein in baculovirus-infected insect cells can bind to B. megaterium and B. thuringiensis but not to Staphylococcus aureus, Escherichia coli or Beauveria bassiana. Consistent with these data, BiPGRP-S shows antibacterial activity against B. megaterium and B. thuringiensis. These results indicate that BiPGRP-S is an inducible protein that may be involved in the immune response against bacterial infection of the genus Bacillus as an amidase-type PGRP-S.

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.

Pseudorabies virus (PRV), a member of the Alphaherpesviridae, is the causative agent of Aujeszky’s disease in pigs. Glycoprotein B (gB) of PRV, a major constituent of the viral envelope, consists of 916 amino acids. We continuously combined three gB epitopes, E1 (aa 62-129), E2 (aa 217-282), and E3 (aa 543-737). The DNA fragment containing the PRV gB epitopes was fused with polyhedrin gene in order to generate recombinant baculovirus that expresses the recombinant polyhedra with PRV gB epitopes under the control of the Bombyx mori nucleopolyhedrovirus polyhedrin promoter. Recombinant baculoviruses were injected into fifth-instar B. mori larvae. SDS-PAGE and Western blot analyses revealed that recombinant polyhedra constitute polyhedrin and PRV gB epitopes, and that the recombinant PRV gB epitopes showed cross-reactivity against antiserum of PRV gB produced from pig. To examine the immunogenicity of recombinant PRV gB epitopes, we injected into mice as model animals. ELISA results indicated that antibody production is increased in a similar manner in the injection of recombinant polyhedra with PRV gB epitopes, either injected recombinant polyhedra as a granule form antigen without adjuvant or injected recombinant polyhedrin as a soluble form antigen with adjuvant. Taken together, these data show that PRV gB epitopes were produced as a granule form antigen by fusing recombinant polyhedra in baculovirus-infected silkworm larvae and displayed the immunogenicity in mice, indicating the efficacy of the granule form antigen as a PRV gB vaccine.

Diamondback moth (Plutella xylostella L.) belonging to genus Lepidoptera is a notorious pest of cruciferous crops worldwide. We evaluated the bioinsecticidal activity of the liquid cultures (LB and NB) of a bacterial strain, Serratia sp. EML-SE1, isolated from a diseased diamondback moth. The pathogenicity of a bacterial strain to diamondback moth was confirmed by the following procedures: treatment of liquid culture on cabbage leaves, ingestion of inoculated cabbage and mortality response. For the test, twenty 3rd instar larvae of diamondback moth were placed on the Chinese cabbage leaf in a round plastic cage (Ø 10 × 6 cm) and sprayed with the liquid cultures. After 72 hours, insecticidal activity of LB and NB cultures of Serratia sp. against P. xylostella larvae showed 91.7% and 88.3%, respectively. In addition, the bioinsecticidal activity on potted cabbage with 14 leaves in a growth cage (165 × 83 × 124 cm) also was similar to that of plastic cage experiment. Summarized, the Serratia sp. EML-SE1 may be a potent candidate as a bioinsecticidal agent to control diamondbac kmoth.

This study was conducted for establishment of Economic injury levels(EILs) of the tea red spider mite, Tetranychus kanzawai on Rubus coreanus. T. kanzawai was innoculated on May 7 in differently 0, 5, 10, 20 and 40 adults per plant. And pesticide was treated in late April, early May, middle May and late May, respectively. After inoculated of T. kanzawai, the density was increased until the mid-June and decreased gradually in all plots. And higher inoculation density were increased higher than lower inoculation density. Growth variables were not different among experimental plots except number of fruit set, but the number of fruit set and yields were decreased with increasing initial mite density. Densities of T. kanzawai on treated time of pesticide was lower in later treatment time than early treatment time. The late April treatments were not effective of pesticide in harvesting season. The rates of yield loss increased with increasing initial mite density. The relationship between initial T. kanzawai densities and yield losses was well described by a linear regression, Y = 0.6545X + 3.0425, R2=0.9313. Based on the relationship, the number of adults per plant which can cause 5% loss of yield was estimated to be approximately 3.0. And EILs was estimated to be approximately 8.3 adults/leaf in mid-May.

Background: Proteolytic enzymes are involved in insect molting and metamorphosis and play a vital role in the programmed cell death of obsolete organs. Here we show the expression profile of cathepsin B in the fat body of the silkworm Bombyx mori during development. We also compared the expression profile of B. mori cathepsins B (BmCatB) and D (BmCatD) in the fat body during the larval-pupal transformation of B. mori in the BmCatB or BmCatD RNA interference (RNAi) process. Results: BmCatB is ecdysone-induced and expressed in the fat body of B. mori during the molting, and the larval-pupal and pupal-adult transformations, and its expression leads to programmed cell death. In particular, BmCatB is highly expressed in the fat body of B. mori during the larval-pupal transformation and BmCatB RNAi treatment resulted in the arrest of the larval-pupal transformation. RNAi-treated BmCatB knock-down sustained the expression of BmCatD during the larval-pupal transformation. On the other hand, BmCatD RNAi up-regulated the expression of BmCatB in the fat body of final instar larvae. Conclusion: Based on these results, we conclude that BmCatB is involved in the programmed cell death of the fat body during B. mori metamorphosis and that BmCatB and BmCatD contribute collaboratively to B. mori metamorphosis

We have determined the complete mitochondrial genome of the yellow-spotted long horned beetle, Psacothea hilaris (Coleoptera: Cerambycidae), an endangered insect species in Korea. The 15,856-bp long P. hilaris mitogenome harbors gene content typical of the animal mitogenome and a gene arrangement identical to the most common type found in insect mitogenomes. As with all other sequenced coleopteran species, the 5-bp long TAGTA motif was also detected in the intergenic space sequence located between tRNASer (UCN) and ND1 of P. hilaris. The 1,190-bp long non-coding A+T-rich region harbors an unusual series of seven identical repeat sequences of 57-bp in length and several stretches of sequences with the potential to form stem-and-loop structures. Furthermore, it contains one tRNAArg-like sequence and one tRNALys-likes equence. Phylogenetic analysis among available coleopteran mitogenomes using the concatenated amino acid sequences of PCGs appear to support the sister group relationship of the suborder Polyphaga to all remaining suborders, including Adephaga, Myxophaga, and Archostemata. Among the two available infraorders in Polyphaga, a monophyletic Cucujiformia was confirmed, with the placement of Cleroidea as the basal lineage for Cucujiformia. On the other hand, the infraorder Elateriformia was not identified as monophyletic, thereby indicating that Scirtoidea and Buprestoidea are the basal lineages for Cucujiformia and the remaining Elateriformia.