• Journal of Sericultural and Entomological Science
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• v.38 no.2
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• pp.118-123
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• 1996

Dietary supplementation of folic acid to silkworm larvae in different concentration (100, 200 and 300 ${\mu}$g/ml) to the 4th and the 5th instar resulted in a significant increase in economic parameters, such as female and male cocoon weights, shell weights, egg productivity and egg hatching percentage. However, larval duration was significantly decreased, as compared with that of carrier control. The increase in the glycogen and protein contents of the fat body was not significant, whereas the increase in trehalose content of haemolymph in all the treated groups and protein content of fat body and haemolymph in 100 ${\mu}$g/ml treated groups were significant. However, the haemolymph protein was significantly decreased in 300${\mu}$g/ml treated group.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.2
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• pp.149-152
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• 2001

Effect of topical application with 10, 20 and 30 ${\mu}g$/ml testosterone propionate and estradiol -17${\beta}$ on the fourth and fifth instar bivoltine NB18 silkworm larvae Bombyx mori, on the glycogen and protein contents of the Fat body and trehalose and protein contents of the haemolymph has been studied. Glycogen content of the fat body was significantly decreased in both testosterone propionate and estradiol -17${\beta}$ treatment groups except in the group treated with 30 ${\mu}g$ testosterone propionate where the increase was not significant when compared with those of carrier controls. The increase/decrease in haemolymph trehalose content did not show any significant difference in all the treated groups. Protein content of the fat body significantly increased in 10 and 20 mg testosterone propionate and estradiol -l7${\beta}$ treated groups but in 30 mg treated groups the increase was not significant when compared with those of carrier controls. There was no significant change in the haemolymph protein content in all the testosterone propionate and estradiol -17${\beta}$ treated groups except in group treated with 10 ${\mu}g$ estradiol -17${\beta}$ where it showed a significant decrease when compared with that of carrier control.

• International Journal of Industrial Entomology and Biomaterials
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• v.36 no.1
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• pp.15-24
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• 2018

N-glycosylation is an important posttranslational modification that results in a variety of biological activities, structural stability, and protein-protein interactions. There are still many mysteries in the structure and function of N-glycans, and detailed elucidation is necessary. Baculovirus expression system (BES) is widely used to produce recombinant glycoproteins, but it is not suitable for clinical use due to differences in N-glycan structure between insects and mammals. It is necessary to develop adequate model glycoproteins for analysis to efficiently alter the insect-type N-glycosylation pathway to human type. The previous research shows the recombinant alpha 1-acid glycoprotein (${\alpha}1AGP$) secreted from silkworm cultured cells or larvae is highly glycosylated and expected to be an excellent research candidate for the glycoprotein analysis expressed by BES. Therefore, we improved the ${\alpha}1AGP$ to be a better model for studying glycosylation. The modified ${\alpha}1AGP$ (${\alpha}1AGP{\Delta}$) recombinant protein was successfully expressed and purified by using BES, however, the expression level in silkworm cultured cells and larvae were lower than that of the ${\alpha}1AGP$. Subsequently, we confirmed the detailed profile of N-glycan on the ${\alpha}1AGP{\Delta}$ by LS/MS analysis the N-glycan structure at each glycosylation site. These results indicated that the recombinant ${\alpha}1AGP{\Delta}$ could be usable as a better model glycoprotein of N-glycosylation research in BES.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.75-80
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• 2006

Changes in the levels of protein and free amino acids in the haemolymph of three selected races of the silkworm, Bombyx mori viz., PM, $NB_4D_2$ and $CSR_2$, were investigated during 4th moult, 5th instar and pupal period. The levels of total protein in the haemolymph, increased from first day of 5th instar till sixth day. From seventh day till spinning, the protein levels decreased in all the three races. A sustained decrease in the haemolymph proteins was observed during the pupal development in all the three races. The levels of free amino acids, which were high during 4th moult, declined through the 5th age of larval development till spinning. PM showed a relatively higher free amino acid level (3.192 mg/ml) in haemolymph followed by $NB_4D_2$ (2.601 mg/ml) and $CSR_2$ (2.35 mg/ml). The free amino acid levels decreased gradually from prepupal stage but increased again at the end of pupal period. Racial differences in the changes in the levels of protein and free amino acids in the haemolymph were observed in the larvae and pupae when subjected to two high temperature regimes of $30^{\circ}C$ and $35^{\circ}C$. The results showed that high temperature induces specific changes in the metabolism (reversible thermal stress) that have different adaptive value in different races of the silkworm. Relatively higher increase in the free amino acid levels in the haemolymph of Pure Mysore presumably provides protective cover to tissues against high temperature by an increase in osmolarity and reduction in evaporative water loss. The absence of such a mechanism may be responsible for temperature susceptibility of the bivoltine races like $NB_4D_2$ and $CSR_2$.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.1
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• pp.15-20
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• 2003

The present investigation was carried out to study the possible cause for reciprocal difference in silkworm hybrids. By utilising the polyvoltine race Pure Mysore (PM) and newly evolved breeds (CSR2, CSR5, CSR16 and CSR17), the direct and reciprocal crosses of polyvoltine ${\times}$ bivoltine and also bivoltine hybrids were studied. The hybrids of polyvoltine ${\times}$ bivoltine (direct) are superior to their reciprocal crosses in respect of cocoon yield, cocoon weight and filament length. The reciprocal crosses of polyvoltine ${\times}$ bivoltine are superior to their direct crosses in respect of fecundity and short larval duration. No significant differences were observed in the characters like cocoon shell ratio, raw silk percentage, denier, reelability and neatness in both polyvoltine ${\times}$ bivoltine direct crosses and their reciprocals. The expression of cocoon characters as a function of sex revealed that direct crosses (polyvoltine ${\times}$ bivoltine) showed higher cocoon weight, pupal weight, shell weight and longer filament length in females than the reciprocal crosses (bivoltine ${\times}$ polyvoltine), where as these characters in males were almost the same in both direct and reciprocal crosses, indicating that the sex-linked genetic factor played a more important role. it was clear that difference in cocoon yield observed in reciprocal crosses of polyvoltine ${\times}$ bivoltine was due to the low cocoon and shell weight in females which was turn due to presence of early maturity genes (Lme) linked with sex-chromosome (X) which effect on larvae period of the silkworm. In bivoltine hybrids, i.e., both direct and their reciprocals crosses, all the characters viz., hatching percentage, larval duration, survival, cocoon weight, cocoon shell weight, cocoon shell ratio, raw silk percentage, filament length, denier, reelability and neatness did not show any significant difference (except number of eggs laid by moth) which could account for presence of same maturity genes (Lm) in both direct and reciprocal crosses. it was clear that reciprocal differences occur when the hybrids are prepared from the parental strains with different voltinism.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.165-173
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• 2003

Cytokines represent an essential part of the innate immune response in mammals. Recently, several studies have reported the presence of cytokine-like activities and molecules in the invertebrates such as echinoderms, tunicates, mollusks and insects. In our serial study, we investigated presence of cytokines in the silkworm, Bombyx mori, infected with several immune inducers. Western blotting analysis using rabbit anti-human cytokines showed the presence of IL-6-like molecule in the hemolymph collected at 8 and 24 hrs after infection with peptidoglycan and oligodeoxynucleotide, and the molecular weight of the proteins was ∼45 kDa. We attempted to isolate the molecule by gel permeation HPLC, anion exchange chromatography, ultra centrifugation, and immuno-dot-blot assay, but until now the effort was not much successful yet. It, however, does not appear that the IL-6-like molecule in the silkworm larvae is a mere experimental artifact happened by Western blotting analysis. Instead, further experiment on this subject probably will provide us more fruitful result as detected in other invertebrates including insects.

• Journal of Sericultural and Entomological Science
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• v.45 no.2
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• pp.108-111
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• 2003

A infectious fungus (J131) recently isolated from artificial cultivation farms of Paecilomyces tenuipes is known to cause a great yield loss and deterioration of products. In this study, its occurrence and characterization were made. The isolate (J131) was identified as Paecilomyces farinosus (Holm ex S. F. Gray) Brown ＆ Smith. Colonies of the isolate on PDA grew moderately fast, attaining a diameter of 19mm within 14 days at 24${\pm}$1$^{\circ}C$. Vegetative hyphae are smooth-walled and hyaline. Conidiophores mainly arose from the submerged mycelium, usually measuring 1.3-1.5$\mu\textrm{m}$ wide and consisted of verticillate branches with whorls of 2 to 4 phialides. Phialides of 3.8-5.8${\times}$1.5-1.8 $\mu\textrm{m}$, with a swollen basal portion tapered into a distinct neck of about 0.6-0.8 $\mu\textrm{m}$ wide. Conidia arisen from the phialides, were ellipsoidal to fusiform, smooth-walled, hyaline, 1.8-2.4${\times}$1.1-1.4$\mu\textrm{m}$. .When silkworm larvae were injected with concentration of 1${\times}$10$\^$8/ conidia/$m\ell$ of infectious fungus J131, infection rate was 41.3％ and synnemata formation rate was 23.6％. Black spots were appeared on the surface of silkworm larvae in 5-9 days after injection. The surface of larvae was covered with hyphae which were gradually turned into yellow from white color during the incubation period of 3-4 days. 1 to several 4-6mm long synnemata of yellow or orange color were formed on the surface. The matured synnemata were ramified from its end and produced much conidia of white color.

• Journal of Sericultural and Entomological Science
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• v.32 no.1
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• pp.31-37
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• 1990

For fine-denier cocoon filament production, the precocious trimolting silkworms were induced by the treatments with both the imidazole compound "KK-42" hating anti-juvenile hormone activity and high temperature, and their growth, dietary efficiency, cocoon and cocoon filament qualities were compared with those of normal tetramolters as control. 1. The percentage of precocious trimolters was higher in the application of KK-42 than the treatment of high temperature. The effective concentration of KK-42 was 10$\mu\textrm{g}$/1arva. The high temperature treatment for 48 hour was more effective than 36 hour treatment, and silkworm larvae fed on mulberry leaves were more sensitive than artificial diet to induce trimolters. 2. The larval duration of the trimolters induced by KK-42 was 5.17 day shorter than that of normal tetramolters in mulberry leaves rearing, and the increasing pattern of body weight in the 4th inster larvae of trimotlers was similar to the 5th instar larvae of normal tetramolters. 3. The qualities such as cocoon weight, cocoon layer weight and cocoon layer ratio of precocious trimolter induced by KK-42 were much lower those of normal tetramolters: the cocoon weight and cocoon layer ratio were 0.78g, 14.2cg and 18.4% in mulberry leaves rearing, and 0.86 cg, 10.3cg and 12.3% in artificial diet rearing, respectively. 4. The size of cocoon and cocoon filament was smaller in the precocious trimolter, both KK-42 and high temperature, as compared with that of the normal tetramolters. 5. The efficiency of cocoon layer production of the precocious trimolters by the KK-42 was lower than that of the normal tetramolters: amount of the cocoon layer production per 1g of dry mulberry leaves ingested was 7.97cg in the precocious trimotors, while 9.20cg in the normal tetramolters.

• Journal of Sericultural and Entomological Science
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• v.47 no.1
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• pp.12-17
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• 2005

To develop technique for the production of P. tenuipes stromata on a large scale, the infection of P. tenuipes and the growth of stroma were investigated by silkworm (Bombyx mori) variety. Also, studied about biological activites of fruiting body formed on silkworm. Infection rate of the 5th instar larvae of the silkworm with P. tenuipes was the highest in Yangwonjam, followed by Hachojam, Baegokjam and Chilbojam in that order. Also, as the inoculation times was increased, infection rate tended to be raised. The rate of fruiting body formation of the silkworm pupae infected with P. tenuipes was the highest in Baegokjam, followed by Yangwonjam and Chilbojam in the order. But, actually the fruiting body formation of the 5th instar larvae of the silkworm tested was good in Chilbojam, followed by Yangwonjam and Baegokjam in that order in 3 times spraying inoculation. The fruiting bodies of Yangwonjam and Chilbojam infected with P. tenuipes had high amount of Mannitol, but Baegokjam and Hachojam had high concentration of Glucose on a dry weight basis. The mean content of total amino acid in the fruiting bodies of P. tenuipes was 1.03 ${\mu}mole/g$. The distribution rate of amino acid components decreased in the order of Arginine (12.2%)>Glycine (10.5%)> Proline (9.6)>Tyrosine (8.9%)>Serine>Leucine>Threonine. The most abundant amino acid in the fruiting bodies of the Baegokjam, Chilbojam and Hachojam infected with P. tenuipes was arginine, while Yangwonjam was Glycine. The most abundant fatty acid in P.tenuipes was Oleic acid on a dry weight basis. The unsaturated fatty acids such as Oleic acid, Linoleic acid and Linolenic acid accounted for more than 78% of the total fatty acids.

• Journal of Sericultural and Entomological Science
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• v.18 no.2
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• pp.83-88
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• 1976

A study was made on the interstrain difference in dose-infection response and induction response of silkworm to the two strains of cytoplasmic polyhedrosis virus to obtain some informations on screening the leading strains. Comparison of dose-infection responses of larvae following the inoculation of the inclusion body of cytoplasmic polyhedrosis virus revealed that there is no significance in the resistance to dose-infection and any of strains were more resistant at advanced stage(4th instar) than younger stage(2nd instar). In the pathogenicity between cytoplasmic polyhedrosis virus with hexagonal and tetragonal outline, the hexagonal polyhedra showed higher pathogenicity than the tetragonal polyhedra. However, the averages of $LC_{50}$ of cytoplasmic polyhedrosis virus with hexagonal outline to larvae of parental inbred line were 6.12${\times}$10$\^$6//$m\ell$ at 2nd instar and 1.57${\times}$10$\^$7//$m\ell$ at 4th instar in spring and those to their hybrids were 1.28${\times}$10$\^$6//$m\ell$ at 2nd ins tar and 4.99${\times}$10$\^$6//$m\ell$ at 4th ins tar in autumn. Meanwhile the $LC_{50}$ averages of tetragonal polyhedra. to larvae of parental inbred line were 2.06${\times}$10$\^$7//$m\ell$ at 2nd instar and 5.67${\times}$10$\^$7//$m\ell$ at 4th instar in spring and those to their hybrids were 9.84${\times}$10$\^$6//$m\ell$ at 2nd instar and 3.86${\times}$10$\^$7//$m\ell$ at 4th instar in autumn, respectively. In comparison of induction response of silkworm larvae to inoculation of tetragonal polyhedra of cytoplasmic polyhedrosis virus, the induction rate of hexagonal polyhedra was remarkably higher in the treatment of inoculation at 2nd instar than at 4th instar and at the concentration of 1.3${\times}$10$\^$6//$m\ell$ than at any others. Most of induction showed a mixed infection with hexagonal and tetragonal polyhedra of cytoplasmic polyhedrosis virus.