• Title/Summary/Keyword: haemolymph

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Change in Major Carbohydrate Contents in Diapausing and Nondiapausing Pupae of the Fall Webworm, Hyphantria cunea (흰불나방(Hyphantria cunea D.)의 휴면용과 비휴면용에서 탄수화물 함량의 변화)

  • Choi, Mann-Yeon;Boo, Kyung-Saeng
    • Korean journal of applied entomology
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    • v.26 no.3 s.72
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    • pp.145-150
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    • 1987
  • Carborhydrate contents were compared between diapausing and non-diapausing pupae of the fall webworm, Hyphantria cunea DRURY. Glycogen content in the whole body of diapausing pupae kept at $-5{\pm}1^{\circ}C$ was less than that of those left at room temperature, while the amounts of trehalose and sorbitol showed the reverse trend. The osmotic pressure of haemolymph was higher in diapausing pupae than that in non-diapausing pupae. The mean oxygen consumption rate of diapausing pupae was $4{\sim}6.7$ times less than that of non-diapausing, normal pupae.

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Synthesis of New Uracil-5-Sulfonamide Derivatives and Immuno-Stimulatory Effect of a Chemically Modified Hemolymph of Biomphalaria alexandrina on Schistosoma mansoni Infected Mice

  • Fathalla, O.A.;Haiba, M.E.;Maghraby, A.S.
    • Archives of Pharmacal Research
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    • v.26 no.5
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    • pp.358-366
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    • 2003
  • Some N-p-substituted phenyl uracil-5-sulphonamide derivatives have been synthesized to be evaluated as molluscicides against Biomphalaria alexandrina snails, the intermediate host of Schistosoma mansoni. Schistosoma mansoni infected mice were treated with hemolymph obtained from pre-treated Biomphalaria alexandrina snails with the products 4a, 10a, 10b and 4b or obtained from non-treated snails. The selection of the concentration based on the predetermined dose which caused mortality of less than 50% of snails/24 h. $LC_{50}$ of compounds 4a, 10a, 10b and 4b was 50, 100, 200 and 50 ppm respectively. The result showed that immuno-stimulatory effect of treated hemolymph with compounds 4a, 10a and 4b was related to significant protective effects (44.4, 34.6 and 50.4% reduction in worm burden respectively). In addition, mean total worm burdens were significantly reduced in non treated hemolymph by 33.8%. The effect of hemolymph obtained from treated or non treated snails on S. mansoni adult worms antigens was studied by indirect immunofluorescence technique using chronic mouse sera (CMS). The results indicated that there was a strong reaction with epitopes in gut epithelium, tubercles, teigument and subtegumental musculature of untreated and treated S. mansoni adult worms antigens. Therefore, treatment of hemolymph obtained from pre-treated snails with compounds 4a, 10a, and 4b can stimulate specific immune response and induce protective effects against S. mansoni infection.

Phenotypic Expression of the Non Molting Gene in The 'Non-molting of Nho' of Bombyx mori (돌연변이 불면잠$(nm^n)$ 의 불면형질의 발현)

  • 유현주;노시갑
    • Journal of Sericultural and Entomological Science
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    • v.35 no.2
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    • pp.93-99
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    • 1993
  • Studies were carried out to investigate physiological and biochemical analysis of recessive lethal mutant "nmn" and to observe the cuticle formation and dermal gland. All nmn homozygous larvae continued to eat a few mulberry leaves, and died without entering into molt. In case of artificial hatching, eggs had a higher nmn individual segregation ratio than in hibernating eggs. The percentage of nmn individuals with the hatching days was alike during 3days period. As a result of histological observation, nmn mutants dermal gland was different from normal dermal gland in form and size. Normal was formed new cuticle in the middle of the molting, but not in nmn mutant. Total protein content in haemolymph of the normal larva was more than that of nmn smutant and there was a difference between normal and nmn mutant protein components.omponents.

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The Effect of Yellow Soil on Mortality of Korean Scallops, Patinopecten yessoensis at Indoor Tank

  • Oh, Bong-Se;Jung, Choon-Koo;Kwon, Mun-Gyeong;Lee, Jung-Sick
    • The Korean Journal of Malacology
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    • v.26 no.3
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    • pp.179-183
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    • 2010
  • In other to understand the effect of yellow soil to mortality of Korean scallops, P. yessoensis, We investigated its mortality at indoor tanks. The environmental conditions such as water temperature, Salinity, Do and pH were continued constantly during the experimental periods. The 100% of survival rate showed in two experiments groups such as 0.1% and 0.4% of concentration of yellow soil and the other groups as 0.05% and 0.2% of concentration of yellow soil was appeared one dead scallop at each group for 8 days of the experiment periods. the gills of scallop in high concentration of yellow soil (0.2% and 0.4% groups) were covered by yellow soil particles so that this group's scallop should be got a high stress from yellow soil. I think this situation will be more continued for long time the scallop will become to dead. The results of bacteriological analysis did not isolated from haemolymph of scallops and no Perkinsus infectious disease in scallops and the scallops showed necrosis and degeneration on digestive grand and gills of scallop.

Purification and Characterization of Arylphorin of the Chinese Oak Silkmoth, Antheraea pernyi

  • Park, Snag-Bong;Kim, Jeong-Wha;Kim, Soohyun;Park, Nam-Sook;Jin, Byung-Rae;Hwang, Jae-Sam;Seong, Su-Il;Lee, Bong-Hee;Park, Eunju
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.1
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    • pp.33-44
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    • 2003
  • The arylphorin was purified from the pupal haemolymph of the Chinese oak silkmoth, Antheraea pernyi, and characterized physiologically and biochemically, The protein was purified by a simple preparative polyacrylamide gel electrophoresis (PAGE) and subsequent diffusive elution. The preparation was shown to be homogeneous by 7.5% native-PAGE. The native molecular weight of arylphorin was 450 kDa with a 80 kDa single subunit, suggesting hexamer, The protein contained high amounts (18.3%) of aromatic amino acids, phenylalanine (9.7%) and tyrosine (8.6%). Therefore, the protein was identified as a kind of a storage protein referred to as an arylphorin. The protein was stained by Schiff's reagent, suggesting a glycoprotein. The protein contained 4.9% (w/w) sugar and mannose and N-acetylglucosamine were major components. Also, degradation of the protein was begun by heat treatment at 90 for 20 minutes. These results showed that the A. pernyi arylphorin in the study is hexamer associated with the six subunits consisting of a 80kDa single subunit, and is different from that of Kajiura et al. (1998) in the subunit composition.

Studies on the Efficiency of the Artificial Diet for the Silkworm, Bombyx mori L. (누에인공사료의 사료효율증진에 관한 연구)

  • 김주읍
    • Journal of Sericultural and Entomological Science
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    • v.28 no.2
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    • pp.38-47
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    • 1986
  • Silkworm(Bombix mori) were reared with modified artificial diets which were mixed with, as additives, leaf powders of Erigeron canadensis L., Cassia tora L., Cyperus anuricus Var.Laxus and Vigna Sinensis NEDL. The effects of additives on silkworm characteristisc of tested plants were summarized as follows ; 1. About 2-5% addition on dry weight base of leaf powders of E. canadensis, C. tora, C. anuricus or V.sinensis to the basic artificial diet promoted feeding response and digestion and resulted in good practical silkworm characteristics. The addition of V.sinensis and C.anuricus showed especially good effects. 2. The syneristic effect between different plant species was not recognized based on the feeding response and digestion of silkworm reared with various combinations of 2-4 different plant additives. 3. Electrophoretic zymograms of estrase, protease and phosphatase on haemolymph, intestine and silkgland were significantly different among treatments. In general, 1 or 2 more electrophoretic bands were detected when feeding response and digestion were promoted. 4. Contents of starch, crude fat, crude protein and inorganic base were apparently higher in the tested plants than in mulberry leaves. However, no volatile ingredent which is directly realted with feeding response was identified.

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Isolation of a New Microsporidian sp. (NIK-5hm) forming Spores within the Haemocytes of Silkworm, B. mori L.

  • Selvakumar T.;Nataraju B.;Chandrasekharan K.;Sharma S. D.;Balavenkatasubbaiah M.;Sudhakara Rao P.;Thiagarajan V.;Dandin S. B.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.1
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    • pp.63-69
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    • 2005
  • While observing silkworm larval samples received from field, microsporidian spores formed within the haemocytes of silkworm haemolymph were observed. The spores of microsporidian sp. were purified and characterized for morphological characters viz., size, shape as well as serological affinity with different Nosema spp. (M$_{11}$ and M$_{12}$). The infectivity of the isolated spores to silkworm was also studied. The microsporidian sp. was found to be highly pathogenic to silkworm, B. mori. The isolated microsporidian sp. was designated as NIK-5hm, which formed ovocylindrical spore in the haemocytes of silkworm and differed in spore size (length, 4.55 $\mu$m & width, 2.10 $\mu$m) and shape from Nosema bombycis (NIK-ls), NIK-2r (Nosema sp. Mysore [3.6 & 2.8 $\mu$m]), NIK-3h (Nosema sp. M$_{11}$ [3.8 & 1.8 $\mu$m]), NIK-4m (Nosema sp. M$_{12}$ [5.0 & 2.1 $\mu$m]) and Lb$_{ms}$ (Nosema sp. in Lamerine breed of silkworm [4.36 & 2.14]). In immonological test (Latex agglutination test), the isolated microsporidian spores did not react with antibody sensitized latex particles of N. bombycis, M$_{11}$, M$_{12}$ and Lb$_{ms}$ and thus are different type of microsporidian sp., parasitic to silkworm, Bombyx mori L.

In vitro Uptake and Accumulation of Purified Storage Proteins into Fat Body Cells from Huphantria cunea Drury (생체밖에서 미국흰불나방 지방세포에 의한 저장단백질의 흡수와 축적에 관하여)

  • Lee, Bong-Hee;Kim, Kwan-Seon;Moon, Myung-Jin;Kim, Woo-Kap
    • Applied Microscopy
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    • v.18 no.2
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    • pp.102-118
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    • 1988
  • This study was carried out to examine in vitro first whether the storage proteins, which the fat bodies of last larvae from Hyphantria cunea secrete into haemolymph, can be uptaked by the fat body cells of prepupa and then how the uptaked storage proteins can be accumulated in the fat body cells, if uptaken. The fat bodies which had been isolated from last instar larvae were cultured in 1 ml of Grace's insect medium containing $50{\mu}l$ of $^{3}H$-leucine (5.0 mCi/mol, Dupont) at $28{\pm}2^{\circ}C$ for 6 hrs. After the homogenates of the cultured fat bodies were centrifuged at 10,000 rpm for 10 minutes, the proteins included in the supernatant were separated by polyacrylamide gel electrophoreses (non-SDS, 6%). The next treatment of the electrophoresed gel was followed by rinsing. A storage protein band of several bands in the rinsed gel was sliced off. With elution of sliced storage protein bands in Tris-glycine buffer, the purification of radioactive storage proteins from fat bodies was finished. After the purified radioactive storage proteins were added in Grace's insect midis containing fat bodies of the prepupae, they were cultured for the randomly following minutes given as 3, 5, 7, 10, 15, 20 and 30 and for the randomly following hours given as 1, 2, 3 and 4 respectively. The double fixations of the cultured fat bodies in aldehyde and $OsO_4$, were followed by preparation of ultrathin sections from Epon-Araldite blocks through dehydration and embedding. The electron microscope autoradiographic treatment of all prepared sections were performed by the dipping method (Kim et al., 1987). The finally prepared specimens were examined with electron microscope. The fat body cells of the prepupa could be found to uptake the storage preteins of the last instar larvae, which were included in the culture medium, mostly by formation of coated vesicles. The in vitro uptake of the storage proteins actively occurred by 30 minutes after the addition of purified storage proteins in the culture medium. After culture for 7 minutes with the storage proteins, the uptaked radioactive storage proteins labelled a number of lysosomal granules. After culture for 20 minutes with the storage proteins, the radioactive storage proteins were finally incorporated and accumulated in lipid droplets and protein granules. The frequency in the fat body cell of radiolabelled lipid droplets occurs approximately 60%, while the frequency, in which the radiolabelled protein granules occurs in a fat body cell, is approximately 40%.

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Effects of 20-hydroxyecdysoneon Adult Development of Diapausing Fall webworm (Hyphantria cunea Drury) Pupae (휴면중인 미국흰불나방 번데기의 성충발육에 미치는 20-hyderoxyecdysone의 영향)

  • Seung Lyeol Gha;Kyung Saeng Boo
    • Korean journal of applied entomology
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    • v.34 no.4
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    • pp.287-294
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    • 1995
  • Studies were carried out to elucidate the efficacy of exogeneous 20-hydroxyecdysone treatment on terminating pupal diapause in the fall webworm. Hyphantria cunea Drury. And the difference was also investigated between normal adult development and post-diapause development after 20-hydroxyecdysone treatment. In the diapause termination rate of pupae treated with 20-hydroxyecdysone after storage for various periods at 16L:8D and $25\pm$$1^{\circ}C$, at the highest rate was observed from the group stored for the longest period and the lowest rate from those stored for 1.5 months. The time needed for adult emergence was inversely proportional to the chilling (at $0\pm$$1^{\circ}C$) period, and the longer its exposure period at low temperature, the higher its sensitivity to 20-hydroxyecdysone treatment. Pupal diapause duration was almost the same, regardless of storage period in the total darkness or at the photoperiod of 16L:8D, and also they successfully emerged to adult even without any experience at low temperature. The oxygen consumption rate in normally developing pupae showed nearly a typical U-form. But, that of diapausing pupae treated with 20-hydroxyecdysone slowly increased and jumped 14 days after the treatment. Pupal diapause began before formation of adult tissues, an a timing of adult tissue formation coincided with ascending timing of the metabolic rate in both normally developing pupae and diapausing pupae treated with 20-hydroxyecdysone. The diapausing pupae treated with 20-hydroxyecdysone was similar to normally developing pupae in band patterns of proteins from haemolymph or fat body.

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Selection of Mutant Silkworm with Oxidation-deficient Haemolymph for Insect Cell Culture (곤충세포 배지 개발을 위한 체액산화지연 돌연변이 누에계통 선발)

  • Choi, Ji-Young;Kim, Jong-Gill;Choi, Young-Cheol;Yoon, Hyung-Joo;Ahn, Mi-Young;Kim, Sam-Eun;Hwang, Seok-Jo
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.47-50
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    • 2007
  • Insect cell culture system has been demonstrated the effective means of producing medical and agricultural products. Furthermore, Fetal bovine serum (FBS) is in wide use in insect cell culture. Silkworm hemolymph was tested to develop as a substitute for FBS and was effective in insect cell growth. Hemolymph is oxidized and darkens visibly during the collection from silkworms due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as $Y_4$, TBO and $wE^b$ showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The absorbance of mutant hemolymph reached the saturation value at $20^{\circ}C$ in each 330 min ($Y_4$), 360 min (TBO) and 450 min ($wE^b$) min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. The cell growth in the medium supplemented with mutant species hemolmph was more effective that in the medium supplemented with Baekokjam species hemolymph.