• Journal of Sericultural and Entomological Science
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• v.26 no.2
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• pp.11-15
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• 1984

To save the labour reguired for separation of copulated moth during egg Production, some of the chemicals available at the market were on screen test for easy separation of copulated moth of silkworm. The obtained results are summarized as following. 1. In a separation ratio of copulated moth along with a treating time, TCTFE(Trichloro-trifluoroethane) completely separated the copulated moth in 10 minutes. The combinations of TCTFE plus Acetone (87.5 : 12.5 V/V) and TCTFE plus Acetic acid (50 : 50 V/V) take 25minutes for the complete separation of copulated moth. Use of Acetic acid solution only makes 100% separation of copulated moth in one hour and foully minutes and non$.$treatment shows only 47.5% of separation in three and half hours. On the other hand. There is no statistical significance between TCTFE Plus Acetone and control in the egg productivity. 2. The combination of TCTFE Plus Acetone (87.5 : 12.5 V/V) does not infuluence the egg productivity of the moth, showing 443 grains for an average number of egg per moth out of which 417 grains are for the number of fertilized eggs while control shows 452 grains for an avarage number of egg/moth and 428 grains for the number of fertilized eggs. However a sing1e use of TCTFE and Acetic acid shows less egg productivity and number of ferilized eggs per moth, respectively as compared to those of the control. In particular, a single use of acetic acid makes an increase of number of non-egg prodncible moth and it seems to be brought due to a chemical damage. 3. In a rearing test of the egg laid by the chemical treated moth, there are no differences among the treatments in all of the useful characters of the larvae; larval duration, survival rate, cocoon yield, single cocoon weight, single cocoon shell weight and cocoon shell ratio. In these regards, it is recognized that TCTFE can be practically used for the separation of copulated moth and the combination of TCTFE and Acetone promote its efiectiveness on the separation of copulated moth.

• International Journal of Industrial Entomology and Biomaterials
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• v.16 no.2
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• pp.37-48
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• 2008

The diapause and non diapause associated proteins of multivoltine silkworm eggs were analysed by two dimensional (2D) gel electrophoresis. The study was made at 0 hr, 24 hrs and 48 hrs after oviposition. A total of four protein spots in diapause eggs at 24 hrs of oviposition and two protein spots in non diapause eggs at 0 hrs of oviposition were observed. All the six protein spots were considered to have association with diapause and non diapause characters. The molecular weight (MW) and isoelectric point (PI) of these 6 protein spots were calculated. The protein spots 1 and 2 observed in 0 hr of non diapause eggs were found to have the MW of 67 and 75 KDa and PI of 8.6 and 8.4 respectively. Similarly the four protein spots observed in diapause egg at 24 hrs of oviposition exhibited MW viz., 15, 17,20 and 25 KDa and PI of 5.3, 5.8, 6.5 and 6.0 respectively. All these 6 identified protein spots were subjected to in-gel digestion and resulted tryptic peptides were analyzed by Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI TOF-MS). Databases searched based on experimentally determined molecular weights of peptides for the determination of the identities of proteins. The identified proteins indicated homology of 34% to 95%. The results indicate that the proteins may playa role in development of diapause and non diapause eggs.

• Journal of Sericultural and Entomological Science
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• v.5
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• pp.25-38
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• 1965

I. Breeding of tussah silkworm(preliminary report). The preliminary examination for bleeding has been carried out since 1963 in tussah silkworms. 1) The strain(l-MG-B)of the heaviest silk quantity was the green silkworm and brown cocoon in univoltine, and the strains(2-G-B, 2-MG-B) of the heaviest silk quantity were also the green silkwom and brown cocoon in bivoltine in both spring and fall in 1965. 2) It looks like the voltinism, the body color and the cocoon color have reached to pure line up to 1965. II. Best place for the winter of tussah pupa. This work was aimed to find out good ways for the winter of tussah pupa. 1) The hatch of bivoltine was better than that of univoltine. 2) The cocoons covered with the leaves were good in the emergence of moth. 3) The cocoons which were kept at natural temperature till the first emergence of moths would show bad in both hatch and emergence. 4) If some of the pupae kept under natural condition were controled at proper temperature for a few days, hatch and laying eggs were best. 5) The best places for the winter were the egg storage and the rearing room. III. Relation between incubation temperature and voltinism. 1) When the tussah pupa are kept at natural temperature during winter, the moths do not come out of the pupa. 2) There is no difference between about 18$^{\circ}C$ and about 25$^{\circ}C$ during incubation in hatching ratio. 3) The tussah silkworms of univoltine in mortality are stronger than that of bivoltine. 4) There is not any relation between voltinism and high or low temperature for pupa and eggs. IV. Induced mutation by gamma-ray and neutron in tussah silkworm. This work was carried out in order to induce the mutation by treating the pupa or the eggs of tussah silkworm with gamma my and neutron. The results obtained are as follows. 1. Though the whole pupa treated with neutron become moths, the moths have no ability to copulate each other. The only moths emerged from pupa treated with neutron, 4000${\gamma}$ are able to lay all un-fertilized eggs, some of which have a hole on the surface and nothing of contents. 2. The non-diapause eggs are treated with neutron in spring, but the hatching ratio is 50∼60 percent, but the whole eggs treated with gamma ray are never hatched. 3. The sensitivity of the pupa to neutron is weaker than that of the eggs. 4. The hatching ratio is in direct proportion to the gamma ray dose. 5. Author find out a new mutant which is excellent in the cocoon quality, so he will do the progeny test next hear.

• Journal of Sericultural and Entomological Science
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• v.13 no.2
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• pp.99-107
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• 1971

In the suboesophageal ganglion of Bombyx mori, the diapause regulator producing cells which may give an information to the diapause factor cells were found by means of electron microscopy. The diapause regulator producing cells had larger granules (2000 to 5000 A$^{\circ}$ in diameter) than did the diapause factor cells which were partially surrounded by the formers. Highly electron-dense material of lysosome in the diapause regulator producing cells was observed in the diapause-egg producer but such lysosomes were not in the non-diapause egg-producer. It was found that many cytoplasmic granules fuse with lysosome, and smaller granules come out of lysosomes. Some implications of the diapause factor cell and the diapause regulator producing cell were discussed.

• International Journal of Industrial Entomology and Biomaterials
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• v.10 no.1
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• pp.19-24
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• 2005

Nutrition is very essential for growth, development and reproduction. The quantitative impetus of nutritional factor on adult characters and reproductive fitness was studied in Antheraea mylitta by providing fresh leaves of Terminalia tomentosa for 1 to 4 times a day to $5^{th}$ instar larvae. All the characters have improved on giving fresh diet more times over the single diet. The adult weight has strong effect on the reproductive potential i.e., female pupa and moth weight, survivability, pupation rate, emergence percentage, fecundity, fertility, egg weight and hatched out larval weight. The improvement recorded to the tune of $70\%,\;77.1\%,\;115.4\%,\;36.7\%,\;45.9\%\;and\;218.3\%$ in larval weight gain, survivability, pupation rate, female pupa weight, moth weight and fecundity respectively when fresh diets were provided 4 times a day against single diet. The adult emergence percentage has increased from $66.4\%\;to\;96.2\%$ and fertility rose from $52.2\%\;to\;93.6\%$. The weight of eggs and the larvae hatched out of it also found to be significantly higher in 4 diets a day condition. The study revealed that optimization of fresh diet frequency should be maintained to obtain higher egg production and fertility in order to get vigorous larvae to continue the generation.

• Journal of Sericultural and Entomological Science
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• v.16 no.1
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• pp.85-92
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• 1974

Many of studies on the transovarial transmission of occult virus and their activation due to various stresses such as cold or heat treatment, chemical feeding, and nutritional deficiency, etc., in the silkworm, Bombyx mori L. have been made, but any attempts have been not made to control virus diseases by detection of the occult virus-carried moths in the production of silkworm egg of hybrids, because of difficulty to detect occult virus in any stage. Therefore, it may be worth while to disclose whether a sublethal infection of the moths from which active virus are detectable, has the same level of induction rate as that of occult virus activation, thus to apply its results for the reduction of the occurence of virus diseases in silkworm rearing. For these purposes, the following experiment was conducted as one of preliminary steps. In this study, investigations on the generation-to-generation transmission of occult virus and a sublethal infection, and the role of chromosomal gene of the host, Jam 103 and Jam 104 in the Previous generation, and Jam 103 x Jam 103 and Jam 104 f Jam 104 in the next generation were made for the induction of virus diseases due to the transmitted virus. The frequency of cytoplasmic polyhedrosis due to the induction in the F$_1$ generation was markedly higher in the cross-batches, male$\times$female and male$\times$female in which inoculated individuals were used as fem ale parents than in the cross-batches, male$\times$female and male$\times$female in which virus has been not inoculated or inoculated only to male in the previous generation. The tendency of increasing rate was observed in any treatments; such as the inoculations of cytoplasmic polyhedrosis virus (10$\^$5/, 10$\^$6/ 10$\^$7, and 10$\^$8//ml ill different concentration of inocula) , cold-treatment (5$^{\circ}C$, 12hrs or 24hrs), and formalin-feeding treatment (2％ or 3％). The shape of polyhedra (tetragonal in outline) examined in the F, larvae was identified as that of the inoculated polyhedra with partial application of immunofluorescent techniques. These results suggests that the cytoplasmic polyhedrosis virus in B. meri L. are transmitted to the next generation through the egg, apparently in the occult state. And the experimental results of various cross-batches revealed the egg cytoplasm plays an important part i the transmission of the occult virus of the cytoplasmic polyhedrosis virus,

• International Journal of Industrial Entomology and Biomaterials
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• v.26 no.1
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• pp.31-40
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• 2013

Paralytic peptide binding proteins (PP-BP) are 30KP proteins that show similarity to ENF binding proteins. The ENF-BP act as active regulators of ENF peptides. ENF peptides are multifunctional insect cytokines. The comparison of gene expression in diapause induced and non-diapause eggs at different time intervals after oviposition showed an upregulation of PP at 18h as well as PP-BP at 12 and 18h after oviposition along with few other genes. The current study has been taken up to investigate the role of PP as well as PP-BP in diapause induction in polyvoltine silkworms and to study the multigene organization of PP-BP in the Bombyx mori genome. The tissue specific expression analysis revealed that, PP-BP is highly expressed in fat body followed by egg and brain while no expression was observed in midgut. The expression levels of PP and PP-BP in diapause and non-diapause eggs from 0h to 48h after oviposition, validated through realtime PCR revealed that PP is highly expressed at 18 and 24h while PP-BP expression is higher at 12 and 18h time intervals suggesting their possible role in diapause induction. The whole genome survey of the PP-BP paralogous sequences revealed a total of 46 B. mori PP-BP homologs that are classified into 3 categories viz., ENF-BP, Typical 30KPs and serine/threonine rich 30KPs. These paralogous sequences are distributed on chromosomes 7, 20, 22 and 24, all 30KP and S/T rich 30KP proteins are present in the same locus of chromosome 20.

• Journal of Sericultural and Entomological Science
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• v.2
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• pp.1-31
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• 1962

The purpose of this treatise is to prove the presence of cystine in silk fiber through wide sampling throughout all the sericultural processes of Bombyx mori.; also to show that disulfide cross linkages exist in the silk fiber. The conclusions reached were as follows: 1. Crystalline cystine was obtained from silk fibroin using Folin's Method. 2. Analytical data showing the cystine content of silk fiber and its related materials were obtained using Sullvan's Method as follows: Material Percent Cystine A. Mulberry leaf protein 0.175 B. Silkworm egg 0.33 C. Silkworm Body, matured, fat extracted, without silk gland 0.41 D. Silk gland, matured 1.23 E. Silkworm feces none F. Silkworm pupa, fat extracted 0.30 G. Silkworm moth, fat extracted 0.60 H. Raw Silk 0.22 I. Fibroin 0.175 J. Sericin 0.30 3. The presence of cystine in the silkworm was substantiated the existence of 0.175 % methionine in mulberry leaves and 0.12% methionine in the silk gland. 4. Part of the sulfhydryl compounds in the silk gland is believed to transfer to serine and methionine, with the former being secreted into the liquid silk finally as silk fiber and the latter used for nutritive purposes in the growing of silk gland tissue. 5. The cystine content is variable by mulberry species, silkworm species, sex, breeding process, and other culturing environments. 6. Hybrid silkworms require more nutritive amino acids for effective growth than the original parents, and secrete less of them as silk fiber. 7. From such an observation, the amino acid composition of silk fiber is believed to be fairly flexible. Cystine if included in the amorphous part of the fiber, especially in sericin. 8. The result from enriching the silkworm diet with pure cystine or wool cystine did not result in any advantage, therefore it is believed that the natural cystine and methionine contents in the mulberry leafaregoodenoughforsilkwormnutrition. 9. The disulfide cross linkage in silk fiber was verified by using the Harris Method. Contraction took place following the treatment of the fiber with various salts and acids. Comparisons were made with wool fiber. 10. During these experiments, the fibrious structure of silk fiber and the net-globular liquid form were photographed microscopically. It is believed that the globules of liquid silk are net-formed by the inter attraction of the OH ion of the globular peptide and the H ion of water as shown by the hair cracking behavior of the film. The net-globular protein precipitation from the mulberry protein solution showed that mulberry is a proper diet for the formation of fibrous protein in the silk fiber. 11. The significance of the presence of cystine in silk fiber as emphasized in this paper should result in modification of the general conception that cystine is absent from this fiber. NOTICE: A part of this treatise was presented at the annual Korea Sericultural Society meeting held in 1961.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.117-121
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• 2004

The patterns of diapause-associated proteins of silkworm eggs were analyzed by two-dimensional (2-D) gel electrophoresis. Among the hundreds of spots on the 2-D gels, at least two proteins were considered to be associated with diapause. A protein, spot 4, with an approximate molecular weight of 38 kDa and pI 6.1 was observed in the HCI-treated, cold-treated, and diapause eggs, respectively. Spot 4 was undetectable in unfertilized eggs and non-diapause eggs at two days after oviposition, suggesting that this protein may be associated with the entrance to diapause. A protein, spot 11, with an approximate molecular weight of 21 kDa and pI of 61 was detected in the unfertilized, HCl-treated, and cold-treated eggs, respectively, after oviposition by normal moths. In diapausing eggs, a protein corresponding to spot 11 was observed in 3-, 5-, and 30-day-old eggs, while the protein was not detected one-day-old eggs. The protein corresponding to spot 11 was not detected in unfertilized and non-diapause eggs obtained from subesophabeal ganglion (SG)-extirpated moths either. Spot 11 was also considered to be a diapause specific protein, which occurred at only early embryonic stage under the control of diapause-downregulated gene.

• Journal of Sericultural and Entomological Science
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• v.36 no.1
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• pp.30-36
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• 1994

choChorion(egg-shell) morphology of the F1 hybrid between Bombyx mori and Bombyx mandarina has been observed by scanning electrom microscope and chorion protein was analyzed by electrophoresis. The chorion surface structure of F1 hybrids in the lateral(flat) region was similar to that of maternal line. The F1 hybrids chorion was found to have basically a three layer structure. The middle and inner layer were very much like those of the Bombyx mandarina and Bombyx mori. There were many conic pillar structures in the outer layer of the F1 hybrid, which was similar to Bombyx mandarina. This conic pillar structure had a thin cover layer was more clear in the dorsal and ventral side of the F1 hybrid chorion. The conic pillar structure of Bombyx mandarina was found to be dominant in F1 hybrid chorion irrespective of their maternal line. Major components of chorion protein were analyzed by two-dimensional electrophoresis and found to have isoelectric points in the range of pH 4.0-6.5 and molecular weight 10 to 50 kd. F1 hybrid chorion protein components related directly to those of the maternal line. The conic pillar structure was dominat characteristic and it was present in all F1 hybrid.