• Journal of Sericultural and Entomological Science
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• v.28 no.1
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• pp.30-36
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• 1986

Polyacrylamide gel electrophoresis and autoradiography were applied to investigate the developmental profiles of the major hemplymph proteins (MHPs) and their biosynthesis. In addition, some biochemical methods were also used to isolate and purify the MHPs. The obtained results are summarized as follows. 1. MHP-a began to appear from the 2nd day of the fourth-instar larva while MHP-b and -c were detected first on the 1st day of the fifth-instar larva. All these proteins, however, showed a drastic increase in concentration at the 2nd day of the fifth-instar larva. 2. MHP-b and -c were synthesized in fat body on early day of the fifth-instar larva, but the possibility of MHP-a synthesis in fat body was excluded. 3. MHP-b was isolated and purified by heat-treatment (6$0^{\circ}C$), gel filtration on Sephadex G-100 and column chromatography on DEAE-cellulose and CM-cellulose. Purified MHP-b showed a single band on polyacrylamide gel- and SDS-polyacrylamide gel electrophoresis.

• International Journal of Industrial Entomology and Biomaterials
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• v.37 no.2
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• pp.95-99
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• 2018

In insect defense system, antimicrobial peptides (AMPs) are one of important biological molecules to survive in a variety of environments. Insect can synthesize AMPs to protect against invading pathogens in humoral immune response. Taking more advantage of biological antimicrobial molecules, we report antibacterial activity of two cecropin type peptides, cecropin and moricin, isolated from the silkworm against four salmonella species. In this work, we purified antimicrobial candidate peptides (AMCP) from the extracts of immune challenged silkworm larval hemolymph by two-step chromatographic purification procedure, cation exchange and gel permeation chromatography. The molecular weights of purified peptides were estimated to be about 4 ~ 5 kDa by Tricin SDS-PAGE analysis, and identified as silkworm cecropin and moricin by NCBI BLAST homology search with their N-terminal amino acid sequences. As antibacterial activity assay, the purified peptides showed stronger antibacterial activity against Salmonella pathogens with an MIC value of $1{\sim}4{\mu}g/mL$. Therefore two cecropin type peptides purified from the silkworm will be valuable potential materials for development of new natural antibiotics.

• The Korean Journal of Malacology
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• v.24 no.1
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• pp.41-50
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• 2008

The histochemical characteristics and ultrastructure of the mantle in the spiny top shell, Batillus cornutus were described using light and electron microscopy. The simple epidermal layer wrapped on the top and bottom of the centrally located connective tissue. And then the epidermal layer were divided into the outer epidermal layer near a shell and the inner epidermal layer closed to the visceral mass. The connective tissue layer was composed of the collagen fiber muscularfiber bundle and hemolymph sinus. Mucous cells in the apical mantle contained acid and neutral mucopolysaccaride, and acidic carboxylated mucopolysaccaride in the mid and marginal mantle. The mantle thickness, epidermal layer thickness and hemolymph sinus area displayed a trend of reduction from the marginal zone to the apical zone. From TEM observation, it was possible to distinguish epithelium, ciliated cell, absorptive cell and secretory cell in the epidermal layer. The epithelia were columnar and the nucleus was elliptical. The free surface were covered with microvilli. The lateral membranes of epithelium was con nected with neighboring cells by the zonular occludens, zonular adherens and membrane interdigitation. Ciliated cell on free surface had cilia and microvilli, and numerous mitochondria in the apical cytoplasm. In the epidermal layer, it observed 2 type cells having absorptive function. The absorptive cells were columnar in shape, and contained microvilli, pinocytotic vesicles, mitochondria and lysosomes of various electron density. Secretory cells can be divided into four types (A, B, C, D) depending on the cell shape and characteristics of secretory granules. These cells were unicellular glands and had similar characteristics to previously reported on the mantle of the gastropod and bivalves.

• Journal of Aquaculture
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• v.21 no.2
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• pp.111-122
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• 2008

This study was conducted to assess the usefulness of the temperature characteristics of the tidal flat sediments during low tide as a environmental factor, and burrowing behaviour, fatness, total hemocyte counts(THCs) and differential hemocyte counts(DHCs) of hard clam Meretrix lusoria as biological activity indices for the management of hard clam farms located in Taean(Chungnam province) and Gimje(Jeonbuk province) tidal flat in Korea. Temperature ranges of the sediment at 1cm depth during spring(March to May) and summer(June to August) in Taean(where the exposure time was about $5.5{\sim}6$ hours during low tide) were $8.7{\sim}26.8^{\circ}C\;and\;27.6{\sim}32.8^{\circ}C$, respectively. Even though there was no significant difference(P>0.05), temperatures of the surface sediment where submerged with remained seawater were generally higher than that of uncovered with seawater. Burrowing depths of normally digged hard clams were 0.9{\sim}3.6cm from March to October, 2002. In the field experiment performed at Taean farming ground covered with seawater, burrowing times of the clams under natural water temperatures were $41.6{\pm}10minutes$ in February and $5.4{\pm}1.3minutes$ in August, respectively, and these were influenced by water temperatures. Fatness of hard clams began to decrease from May(at Taean tidal flat) and June(at Gimje tidal flat), showed the lowest level in August and increased again from September. Total hemocytes counts in the hemolymph of the hard clams were decreased to the lowest level in July($24.7{\times}10^4cells/mL$, at Taean tidal flat) and August($28.2{\times}10^4cells/mL$, at Gimje tidal flat), and significantly increased again from September(at Taean tidal flat, P<0.01) and October(at Gimje tidal flat, P<0.001), respectively. We observed three types of hemocytes from the hemolymph of hard clams according to whether hemocytes retain the granules or not and the size of the granules. As a results, we could found that periodical monitoring of the sediment temperatures, clam burrowing behaviour and hemocyte parameters were very helpful for the management of hard clam farming.

• Journal of fish pathology
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• v.22 no.3
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• pp.293-303
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• 2009

Effects of stress on the low salinity stress were examined in the pacific abalone Haliotis discus discus. Changes in survival rate, hemolymph count, antioxidant enzyme activities (catalase: CAT and superoxide dismutase: SOD), respiratory burst activity, phenoloxidase activity, lysozyme activity and expression of heat shock protein 70 (HSP70) mRNA were measured 0, 3, 6, 12, 24 or 48hours after low salinity treatment with 25, 30, 33 and 35 psu. Survival rates of pacific abalone were 100% at 33 and 35 psu, but 93 and 97% at 25 and 30 psu for 48 hours, respectively. Hemolymph counts decreased in the time elapsed-dependent way at all of the experimental groups. At low salinity, 25 and 30 psu, SOD and CAT activity increased compared to the experimental group of 33 psu. Moreover, respiratory burst activities of the pacific abalone seemed to have no effect on low salinity stress at any experimental group. However, phenoloxidase activity is an important component of the defence against pathogen that was decreased in a reduction of salinity dependent way. Lysozyme activity also immediately reduced at 25 psu experimental group for 48 h. The HSP70 mRNA was weakly expressed at 33 psu, but strongly detectable at 25 psu experimental group. The HSP 70 mRNA expression in gill increased in the time elapsed-dependent way at 25 psu experimental group and then recovered at 48 h. These results suggest that low salinity stress give rise to inhibitory action of immune system as a result of the decrease of phenoloxidase and lysozyme activity in the pacific abalone, especially.

• The Korean Journal of Zoology
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• v.32 no.4
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• pp.337-350
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• 1989

In this study, we compared patterns of hemolvmph with ovarian proteins during the yolk formation of Gewis paludum by using SDS/polyacrylamide gel electrophoresis and he dimen-tlonal electrophoresis. In addition, we examined patterns of glycoprotein which were composed of yolk substances. The results were as follows: The protein patterns in ovary of the 5th instar without eggs were similar to those of adult after ovulation. Protein amounts of the ovaw without developing eggs were less than those of the ovaw containing oocvtes or matured eggs at the molecular weights range from 66, 000 to 110.000 daltons. No glvcoproteins were observed in ovaw without eggs. But the glvcoproteins were gradually increased according to development of eggs in the 5th instar. After the ovulated ovaw of adult, no glvcoprotein bands urere occurred as the bands of the ovary without eggs in the 5th instar. Also, amounts of hemolvmph protein between 66, 000 and 110, 000 in molecular weight were increased during yolk formation of the 5th instal. The results suggest that ovarian protein substances may originate from hemolpnph. In the 5th instar, the amounts of glycoprotein of developing eggs was gradually increased in the hemolymph. The band of M.W. 29, 500 was occurred in the hemolpnph of the 5th instar and adult without eggs. This protein mal be precursor of glvcoprotein in the high molecular weight area in the ovary of more developed eggs. The number of spots and the amounts of protein in ovary without eggs were less than those of ovary containing eggs by two dimensional elec-trophoresis. The protein bands between 45, 000 and 110, 000 almost appeared in acidic field of he dimensional gel. Especially, the band, M.W. 109, 000, uras separated 3 spots, a1, a2, and as. The band, M.W. 102, 000, has spots which designated b1, b2, b3, and b4. Besides, proteins below M.W. 24, 000 were occurred less spots in the basic field than those of acidic field. The mechanism of intracellular organs (= cell organells) uras related to the yolk protein synthesis of oocyte in the process of yolk protein formation of derris pcfudum.

• Korean journal of applied entomology
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• v.59 no.3
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• pp.185-202
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• 2020

The diapause induction season in Ostrinia furnacalis (Lepidoptera: Crambidae) was estimated in Suwon. Three batches of adult generations were observed, the first one from early May to early July, the second from early or mid-July to early or mid-August, and the third from mid-August to October. In outdoor larval rearing, colony rearing occurring from mid-July to mid-August produced both non-overwintering and overwintering larvae, whereas late-reared colonies produced only overwintering larvae. Larvae collected during July and August in maize fields produced both non-overwintering and overwintering larvae, whereas late-collected larvae produced only overwintering larvae. The results indicated that O. furnacalis has a bi- or trivoltine complex life cycle in this area. In the laboratory, when larvae of all instars within 9 h after molting were first treated to a diapause induction condition (11:13 h = light:dark photoperiod and 20℃), almost all larvae were induced to diapause. However, when similar treatments were conducted age-specifically for the 5th instar larvae, diapause induction rates in 3- and 4-day-old larvae of the 5th instar decreased. In contrast, when larvae were subjected to the diapause induction treatment only during the periods from the hatching stage to the 2nd, 3rd, and 4th instar, almost all larvae were not induced to diapause. The results suggest that the early age of the 5th larval instar is the last stage for sensitivity to diapause induction stimuli. In the diapause-induced larvae, hemolymph trehalose content increased and body supercooling points dropped, compared with those in non-diapause larvae.

• Korean journal of applied entomology
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• v.56 no.2
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• pp.147-152
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• 2017

Heart contraction is essential for insect hemolymph circulation, in which various physiological changes need control of the heart contraction. Thus, interruption of heartbeats intimidate insect survival. Chlorine dioxide induces release of reactive oxygen species (ROS), which has been regarded as the main insecticidal agent. This study analyzed the effect of chlorine dioxide on heartbeats of the Indianmeal moth, Plodia interpunctella. The larvae have the dorsal vessel on the medial dorsal region in a form of an extending tube from the first thorax to 10th abdominal segment. Dorsal vessel contraction was noticeable between 3rd and 10th abdominal segments, where five heart chambers were observed. Average heartbeat rate was 118.6 beats per min at $25^{\circ}C$. However, the heartbeats varied with ambient temperature. Injection of chlorine dioxide to hemocoel suppressed the heartbeats in a dose-dependent manner. The suppressive effect of chlorine dioxide also increased with exposure time to gas form of the chemical at 100 ppm. However, vitamin E (an antioxidant against ROS) treatment significantly rescued the larvae treated with chlorine dioxide in the heart contraction activity. These results suggest that chlorine dioxide negatively influences on the heart contraction of P. interpunctella via its production of ROS.

• Applied Microscopy
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• v.33 no.3
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• pp.223-231
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• 2003

Histology and ultrastructure of the gill in the granular ark, Tegillarca granosa are described using light and transmission electron microscopy. The gill of the clam have typical structure of the filibranch type. The gill filament have several band of lateral and apical cilia. The epithelial layer surrounding the hemolymph sinus is simple and consists of epithelial cells, ciliated cells and secretory cells. The epithelial cells are usually squamous and covered with microvilli. The ciliated cells are usually columnar and can be divided into two types (A and B). Type A cells are more abundance and have lower electron density than B cells. Ultrastructure of the cilia showed that '9+2' microtubular structure of the axial filament and '$2{\times}9$' proximal centriole structure in the cross section. Secretory cells are mainly observed in the apical region of the filament and can be divided into three types of A, B and C with morphological features of the secretory granules. Type A cells of oval shaped are more abundance than other secretory cells and contains numerous secretory granules of low electron dense. Type B cells contains secretory granules of membrane-bounded and high electron dense. Secretory granules of type C cells are elliptical and fine granules surrounding the homogeneous core materials.

• Journal of Sericultural and Entomological Science
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• v.27 no.2
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• pp.20-27
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• 1985

This study was carried out to investigate the histological changes after the infection of nuclear and cytoplasmic polyhedrosis viruses(NPV, CPV) and the resistance to the viruses in various varieties of the silkworm fed on artificial diet. The results obtained were as follows; Among four varieties of silkworm tested, Jam 107$\times$Jam 108 was more resistant than the other varieties tested and Jam 119$\times$Jam 120 was the most resistant to CPV. In case of peroral infection with NPV, Jam 107$\times$Jam 108 showed lower mortality than the remained varieties in low concentration (104/ml). However, all varieties showed high mortality as the concentration of viruses was increased. With infection of CPV, the varieties showed high mortality at the concentration of 107 and 108/ml, while Jam 119$\times$Jam 120 showed the lowest mortality at virus concentration of 104/ml. The fat bodies, epidermal cells and tracheal epithelial cells showed high susceptibility to NPV to break the cells completely and liberate the debris to the body cavity. The CPV infected only the cylindrical cells of mid-gut and formed polyhedrons. In some cells, CPV was liberated to gastral cavity. In the electrophoretic pattern of hemolymph protein of silkworm larvae infected with NPV, bands were dimmed and disappeared as symptom aggravated after infection. Electrophoretic pattern of homolymph proteins of silkworm larvae infected with CPV showed no numerical difference at the later stage of infection, and one or two bands was observed along with lowering the concentrations.