• International Journal of Industrial Entomology and Biomaterials
• /
• v.23 no.2
• /
• pp.231-238
• /
• 2011

Attacin is an insect antibacterial protein that plays an important role in immune response to injury and infection. In this report, we have isolated and characterized of cDNA encoding for the attacin from the immunized larvae of swallowtail butterfly, $Papilio$ $xuthus$. A full length cDNA of $P.$ $xuthus$ attacin was obtained by employing annealing control primer (ACP)-based differential display PCR and 5' RACE. The complete $P.$ $xuthus$ attacin cDNA was comprised of 949 bp encoding a 250 amino acid precursor. It contains a putative 18 amino acid signal peptide sequence, a 42 amino acid propeptide sequence, and a 190 amino acid mature protein with a theoretical molecular mass of 19904.01 and a pI of 9.13. The putative mature protein of $P.$ $xuthus$ attacin showed 48-52% and 24-30% identity in amino acid sequences with that of lepidopteran and dipteran insects, respectively. Semiquantitive RT-PCR results revealed that the transcript of $P.$ $xuthus$ attacin gene was up-regulated at significant levels after injection with bacterial lipopolysaccharide (LPS). We sub-cloned cDNA fragment encoding mature $P.$ $xuthus$ attacin into the expression vector, highly expressed in $E.$ $coli$ BL21 cells, and its antibacterial activity was analyzed. Recombinant $P.$ $xuthus$ attacin evidenced considerably antibacterial activity against Gram-negative bacteria, $E.$ $coli$ ML 35 and $Klebsiella$ $pneumonia$.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.28 no.1
• /
• pp.1-4
• /
• 2014

The effect of an artificial diet on developmental rate, a life history parameter, was examined for the swallowtail butterfly Papilio xuthus. Artificial insect diets are an essential component of many insect rearing systems that produce insects for research purposes. Complex agar-gelled diets are generally prepared in large batches and used shortly after preparation because the degradation of perishable diet ingredients, such as vitamins and fatty acids, can adversely affect insect quality (Brewer 1984). However, the timing of diet preparation may be inconvenient, and large batches wasteful, if the unused excess is discarded. The percentage of pupation varied considerably, with no significant differences among diets, on which a maximum pupation percentage of 83% was observed. Pellet-type diets were investigated with the aim of developing a more easily prepared diet. The extrusion of the artificial diet under high temperature and pressure may induce desirable chemical and physical changes in the extruded product. The purpose of the present study was to develop an artificial diet for rearing P. xuthus.

• Food Science and Preservation
• /
• v.17 no.5
• /
• pp.752-756
• /
• 2010

Lepidoptera (butterflies) extracts, traditionally employed as medicines, have various biological activities. Five species of Lepidoptera (Papilio maackii, Papilio xuthus, Pieris rapae, Eurema hecabe, and Sasakia charonda) were extracted with distilled water (DW), dimethyl sulfoxide (DMSO), ethanol (EtOH), and methanol (MeOH). Each extract was analyzed for anti-oxidant and anti-inflammatory activities using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay method, the ferric reducing ability of plasma (FRAP) test, and a cyclooxygenase-2 (COX-2) promoter assay. The results suggest that Lepidoptera extracts have valuable anti-oxidant and anti-inflammatory properties, supporting the idea that the extracts may serve as a food biomaterial(s) preventing oxidative processes and inflammatory damage.

• Journal of Life Science
• /
• v.17 no.8 s.88
• /
• pp.1046-1052
• /
• 2007

Transferrin is a molecule carrying iron to store and maintain for iron homeostasis of living organisms. In this study, we have purified transferrin, as an iron-binding protein, from the last larval haemolymph of Papilio xuthus by KBr density gradient ultracentrifugation and gel filtration (superose 6 HR) using fast protein liquid chromatography (FPLC) and transferrin containing iron was identified by Ferene S staining. The purified haemolymph transferrin was shown to have molecular mass of 78 and 80 kDa and amino acid composition of transferrin was rich in aspartic acid, valine, leucine and glutamic acid. With immuno-diffusion assay, we confirmed the existence of the transferrin in the haemo-lymph and fat body by detection of visible and clear positive reaction. From the quantitative comparison by rocket immuno-electrophoresis process, the amount of transferrin were increased in the haemolymph of 3 days after pupation and the whole 5 days after pupation. Here, with biochemical and immunohistochemical analysis, we speculate the relationship of transferrin between the physical characteristics and distribution during metamorphosis of P. xuthus.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.31 no.2
• /
• pp.79-84
• /
• 2015

A new cecropin-like antimicrobial peptide (Px-CLP) gene was isolated from the immunechallenged larvae of the swallowtail butterfly, Papilio xuthus, by employing annealing control primer (ACP)-based GeneFishing PCR. The full-length cDNA of Px-CLP is 310 nucleotides encoding a 70 amino acid precursor that contains a putative 22-residue signal peptide, a 4-residue propeptide, a presumed 37-residue mature peptide, and an uncommon 7-residue acidic pro-region at the C-terminus. The deduced amino acid sequence of Px-CLP showed significant identities with other Lepidopteran cecropin D type peptides. RT-PCR revealed that the Px-CLP transcript was detected at significant level after injection with bacterial lipopolysaccharide (LPS). The peptides with or without C-terminal acidic sequence region were synthesized on-solid phage and submitted to antibacterial activity assay. The synthetic 37-mer peptide (Px-CLPa), which removed C-terminal acidic sequence region, was showed exclusively antibacterial activity against E. coli ML35; meanwhile, a 44-mer peptide (Px-CLPb) with C-terminal acidic peptide region was not active. This result suggests that Px-CLP is produced as a larger precursor containing a C-terminal pro-region that is subsequently removed by C-terminal modification.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.8
• /
• pp.1185-1188
• /
• 2010

Previously, papiliocin was isolated from the swallowtail butterfly Papilio xuthus and its antimicrobial activity was suggested. In this study, the antifungal mechanism of papiliocin against Candida albicans was investigated. Confocal laser scanning microscopy (CLSM) and 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence analysis indicated that papiliocin disturbed the fungal plasma membrane. Moreover, the assessment of the release of FITC-dextran (FD) from liposomes further demonstrated that the antifungal mechanism of papiliocin could have originated from the pore-forming action and that the radius of the pores was presumed to be anywhere from 2.3 to 3.3 nm.

• Journal of Life Science
• /
• v.19 no.2
• /
• pp.228-233
• /
• 2009

The purpose of this study is to evaluate the activities of five different antioxidant enzymes in various tissues of Papilio xuthus during pupal stage. Superoxide dismutase (SOD) activity in haemolymph was the highest just after pupation and then decreased gradually until 7 days after pupation but the activity in other tissue was constant during metamorphosis. This result indicates that primary antioxidant process of reactive oxygen species proceed in haemolymph. Catalase (CAT) activity in studied tissues was also the highest just after pupation and its relative activity was also high during pupal stage, suggesting that CAT is the primary enzyme in catalysis of hydrogen peroxide. Glutathion peroxidase (GPX) activity was constant and its relative activity was very low in all tested tissues. Glutathione S-transferase (GST) activity in haemolymph was high at 3 days and 5 days after pupation, and the activity in fat body was the highest at the 1 day after pupation and then decreased gradually for 7 days after pupation. Glutathion reductase (GR) activity in haemolymph and fat body was high at 1 day after pupation, but relatively low GR activity was detected in the rest tissues. Based on these results, GST activity was higher than that of GPX and GR, and it is also believed that GST was more involved in reduction process through lipid peroxidation than GPX.

• Journal of Korean Society of Rural Planning
• /
• v.21 no.4
• /
• pp.1-12
• /
• 2015

Various types of parks such as ecological park, dragonfly park, etc. are tried to design in order to resolve different environmental issues in urban areas. Parks are considered organism habitats that not only improve the biodiversity but also give people a chance to observe living organisms in urban area. This study was conducted to select basic materials for planning a butterfly garden through reference reviews. The following 21 species of butterflies belonged to 1 order, 4 families, and 16 genera were selected: Phengaris teleius, Pseudozizeeria maha, Thecla betulae, Argynnis hyperbius, Dichorragia nesimachus, Hestina assimilis, Polygonia caureum, Sasakia charonda, Vanessa cardui, Vanessa indica, Boloria selene, Byasa alcinous, Papilio machaon, Papilio macilentus, Papilio protenor, Papilio xuthus, Papilio bianor, Sericinus montela, Colias erate, Eurema hecabe, and Pieris rapae. 85 species belonged to 22 families, 59 genera, 74 species, 10 varieties, and 1 subspecies were selected for host and nectar plants for butterflies selected above. These results would be expected to useful in butterfly gardening. After that, we think it is necessary to be applied these study results in field. This study requires a test in the butterfly garden. The test results will be make the butterfly-plant matrix.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.9
• /
• pp.1305-1309
• /
• 2020

Insects possess biological defense systems that can effectively combat the invasion of external microorganisms and viruses, thereby supporting their survival in diverse environments. Antimicrobial peptides (AMPs) represent a fast-acting weapon against invading pathogens, including various bacterial or fungal strains. A 37-residue antimicrobial peptide, papiliocin, derived from the swallowtail butterfly Papilio xuthus larvae, showed significant antimicrobial activities against several human pathogenic bacterial and fungal strains. Jelleines, isolated as novel antibacterial peptides from the Royal Jelly (RJ) of bees, exhibit broad-spectrum protection against microbial infections. In this study, we developed a novel antimicrobial peptide, PAJE (RWKIFKKPFKISIHL-NH₂), which is a hybrid peptide prepared by combining 1-7 amino acid residues (RWKIFKK-NH₂) of papiliocin and 1-8 amino acid residues (PFKISIHL-NH₂) of Jelleine-1 to alter length, charge distribution, net charge, volume, amphipaticity, and improve bacterial membrane interactions. This novel peptide exhibited increased hydrophobicity and net positive charge for binding effectively to the negatively charged membrane. PAJE demonstrated antimicrobial activity against both gram-negative and gram-positive bacteria, with very low toxicity to eukaryotic cells and an inexpensive process of synthesis. Collectively, these findings suggest that this novel peptide possesses great potential as an antimicrobial agent.

• Journal of Sericultural and Entomological Science
• /
• v.51 no.2
• /
• pp.173-179
• /
• 2013

This study was investigation for primary bionomical characteristics, emergence rate of wintering type pupa and oviposition preference, of Papilio xuthus. The emergence rate of wintering type pupa stored at cold chamber($4{\pm}1^{\circ}C$) was 85%, after 10 months. The average emergence rate of wintering type pupa during storage was 89.6%, and the emergence periods were $7.9{\pm}1.9days$ in room temperature. Mating ratio in large oviposition room, $6,000{\times}6,000{\times}3,500$ mm($W{\times}L{\times}H$), was $86.7{\pm}5.8%$ higher than $63.3{\pm}15.3%$ in small room, $2,500{\times}3,000{\times}2,000$ mm($W{\times}L{\times}H$). But the number of eggs in the large room were $137.0{\pm}16.5$ fewer than $141.7{\pm}20.4$ in the small room. As a result of oviposition preference experiment on three different host plants, 10 mated female laid $141.7{\pm}27.8$ on the leaves of Phellodendron amurensew more than $67.7{\pm}20.6$ on Zanthoxylum schinifolium and $77.0{\pm}21.8$ on Citrus unshiu. The egg periods were $4.4{\pm}0.8$ days after oviposition, and the hatchability was 92.2% in $25^{\circ}C$ room condition. The larval period were $19.9{\pm}2.1$ days in the high temperature, long day condition($25^{\circ}C$, 14L : 10D). The headwidth of each developmental larval stage were $0.72{\pm}0.02$(1st), $1.19{\pm}0.02$(2nd), $1.65{\pm}0.05$(3rd), $2.43{\pm}0.07$(4th), $3.21{\pm}0.12$(5th). The pupal ratio was 91.6%. The pupal period were $8.8{\pm}0.9$ days, and the emergence rate was 92.2%.