• Journal of Sericultural and Entomological Science
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• v.26 no.2
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• pp.1-6
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• 1984

The present study was made to establish the purification method of mature microsporidian spores by iso-density equilibrium technique using percoll and the serological discrimination by an indirect fluorescent antibody technique. The purification of new microsporidian spores took effect in the three steps purification method(precentrifugation-percoll iso-density equilibrium centrifugation-rising). There were clear differences in the size of spores between the new microsporidia and N. bombycis. The spores of N. bombycis is short elliptical of 2.07 in a ratio of length to width in diameter while that of new microsporidia is characterized with long elliptical shape which shows a ratio of 2.76 in length to width in diameter. The specific antigens of new microsporidia K79 spores was showed in the spores wall by the indirect fluorescent antibody reaction, and it was affected by the antibody against N. bombycis which antiserum was diluted in 1:20. It means that the new microsporidia K79 is serologically not identical to N. bombycis.

• Journal of Sericultural and Entomological Science
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• v.7
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• pp.49-52
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• 1967

In the F$_1$ hybrids and parents of silkworms, the silkworms of 47 blocks in summer and 68 blocks in fall in 1965 were reared in order to know the effect of mortality on the difference of sex ratio. The results analyzed are as follows; 1. In Moran of Japanese strain-parent and Morean x Daedong of F$_1$ hybrid, there is of no effect of an increase and decrease of mortality on the sex ratio. So male is not always stronger than female. 2. In Daedong of Chinese strain-parent, the more the mortality in the blocks is, the higher the survival ratio of male pupa becomes. (Y=0.233x-41.67). 3. As the results mentioned above, the present investigation method of cocoon quality of Chinese strains with the same number of cocoons of both sexes will have to be considered again, according to the cocoon crops, as far as concerned with the reasonable sampling from the population.

• Journal of Sericultural and Entomological Science
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• v.42 no.1
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• pp.58-64
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• 2000

This study was designed to investigate the effects of silk fibroin powder (Mw 500) on oxidative stress and membrane fluidity in liver membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10g) were fed basic diet (control group), and experimental diets (SEP-2.5 and SFP-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. Cholesterol levels resulted in a significant decrease (12.1% and 9.0%, respectively) in the liver mitochondria and microsomes of SEP-5.0 group compared with control group. Membrane fluidity as significantly increased (16.1% and 16.5%, 5.8% and 17.4%) in the liver mitochondria and microsomes were significantly inhibited (16.1% and 18.3%, 8.1% and 15.1%, respectively) at the SFP-2.5 and SEP-5.0 groups compared with control group. Induced oxygen radicals (BOR) in liver mitochondria and microsomes were significantly inhibited (16.1% and 18.3%, 8.1% and 15.1%, respectively) at the SFP-2.5 and SEP-5.0 groups compared with control group. Induced oxygen radicals (IOR) in liver microsomes were significantly inhibited (17.0% and 26.6%, respectively) at the SFP-2.5 and SFP-5.0 groups compared with control group, but IOR in liver mitochondria was significantly inhibited about 12.3% at the SWP-400 group only compared with control group. Lipid peroxide (LPO) levels were significantly decreased (8.3% and 18.0%, 13.4% and 18.4%, respectively) in the liver mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were dose-dependently decreased (5.4% and 11.6%, 19.0% and 24.4%, respaectively) in the iver mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. These results suggest that administration of SFP may play an effective role in attenuating an oxidative stress and increasing a membrane fluidity in liver membranes.

• Journal of Sericultural and Entomological Science
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• v.1
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• pp.15-21
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• 1960

4231년 농림부산림국에서 송충구제책으로 경기도 임업시험장에서 인공배양한 No.7호균외 수종을 사용할 계획인바 비에 대한 임업시험장 시험보고에 의하면 핵균은 송충에 대하여 맹독성이지마는 잠아에 대하여서는 기독성이 경미하다는 보고 이였으므로 양잠가에 대한 의문을 풀기 위하여 송충 및 잠아에 대한 전염성의 강약을 비교 시험하여 그 독성정도를 구명코저 함. 1. 경기도 임업시험장 연구보고 제1호에 기재된 No.7 호균의 시험성적과 본시험성적과를 대조하면 그 이병률은 일정한 경향을 표시치 않으며 다소 상치된 결과이었다. 즉 별표에 의하면 송충은 72~100% 잠아에 있어서는 4289년도에 0~2% 4290년도이 0~20%였으나 본시험결과 (표 1,2,3)는 송충은 표준구에 6.6% 처리구에 13.3~20%이고 잠아는 5~50%의 이병률이였다. 따라서 송충에는 그 감염도가 높으며 잠아는 그 감염도자 낮다고하는 근거를 인정키 난하다. 2. 본시험에 공시된 접종균 No.7호균은 경기도 임업시험장에서 호칭하는 황황강균(Insarica farinosa)이 었으나 핵균 배양시에 이균이 침입된 관계인지는 모르나 본시험에서 폐사를 검정한 결과 백강균(Botrytis Bassiana)이였다. 따라서 No.7호균에 대한 순도가 의심되며 시험결과의 정확을 기할수가없었다. 3. 경기도 임업시험장연구보고 제1호에 접수된 양년간시검성적에 의하면 경화병균 28종중 가장 이병률이 낮다고하는 No.7호균이 본시험결과에서 잠아에 5~50%의 이병률을 나타내는것으로 보아 소위 경기도 임업시험장에서 순수분리한 균은 어느것이라도 송충에 접종하여 산지에 산포하면 그 균이 중간기생 (인교목, 직교목)을 통하여 야생화되어서 더욱 잠아에 대한 전염력을 강렬하게됨으로 양잠상 위험할것이다. 일본의 소산씨 문헌(경기도 임업시험장도서관)에 의하면 야생혼충에서 전염되는 경화병균의 독성은 잠아에 대하여 강력하다는 점과 $\ulcorner$잠업지조선$\lrcorner$제11호에 전라남도 원잠종제조소 강본태대랑씨의 시험성적(참고문헌)이 이를 여실히 입증하는 바이다. (중략)

• Journal of Sericultural and Entomological Science
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• v.23 no.2
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• pp.37-49
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• 1982

Larvae of the silkworm (Bombyx mori L.) were reared during the 5th instar on the four kinds of artificial diets on the basis of the different amounts of soybean meal used as the protein source. In this experiment it was shown that the various levels of protein in the diet affected not only the growth and silk production but the digestibility of the diet. haemolymph protein and uric acid excretion. The results obtained are summarized as follows; 1. By an increase of the level of protein in the diet the apparant digestibility was increased. but the protein digestibility was comparatively decreased. 2. Larval body weight increment was not observed by the 3rd day of the 5th instar, but was increased from the 4th day as the level of protein was increased in the diet. 3. After the 3rd day of the 5th instar, protein content in the hemolymph was increased steeply by an increase of the protein content in the diet. However, the percentage of hemolymph protein to the ingested protein was decreased from the 2nd day of the 5th instar and increased more or less after the 4th day. 4. An increase of the uric acid excretion was observed as the content of protein in the diet was increased but the pattern of the uric acid excretion was different between high and low-protein diet. However, the percentage of the uric acid excretion to the ingested protein and to the hemolymph protein were both decreased steeply after the 2nd day of the 5th instar. 5. It was also evident that the high-protein diet increased the cocoon productivity. 6. It showed that the feed efficiency for body weight increment and silk formation was high by an increase of the level of protein in the diet, but the protein efficiency was not.

• Journal of Sericultural and Entomological Science
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• v.13 no.1
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• pp.23-29
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• 1971

As a result of analyzing the change of material substance of all sorts biochemically and comparing the control with ${\gamma}$-ray irradiation (800${\gamma}$, 40 Min), incubating the silkworm eggs (Bombyx mori L.) as the objective in the process of growth of embyro shortly before hatching, the following conclusion has been found. 1. Ascorbic acid has shown the maximum increase of 319 r/g in the Byong B embyro stage and in other words it has increased during the period of vigorous metabolism of the materials in eggs but it has decreased before hatching after that period. 2. Triglyceride has shown the increase of 27.54 mg/g in the Byong A stage, the early period of incubation and in other words it has increased in the period of activation of cells in eggs but it has gradually decreased during the growth of embyro after that period. Great change of either total cholesterol or free cholesterol has not been shown from the early period till shortly before hatching. 3. Free fatty acid has shown the minimum decrease of 257.4$\mu$ mole/g in the Byong A stage in which triglyceride increases greatly. On the contrary, it has shown the increase of 1, 020.0$\mu$ mole/g in Ki A stage in which triglyceride decreases. As a whole, the fact that free fatty acid increases according to the growth of embyro in eggs has been found. 4. Glucose has shown the increase of 281.2 mg/g in control during tile Pigment stage and it has shown the increase of 179.6 mg/g in ${\gamma}$-ray irradiation during the same period. The difference in quantity between the former and the latter is due to the fact that the growth of embyro has been influenced by the radio active. Glucose has changed with free fatty acid and phosphorus the other way round. 5. Control organic phosphorus has shewh the increase of 5.23mg/g during the Byong B or Ki A in which organ and tissue in the emhyro has been formed. Organic phosphorus in ${\gamma}$-ray irradiation has shown the increase of 5.73mg/g during Ki B. Inorganic phosphorus has shown only a little change in the control and ${\gamma}$-ray irradiation. The phosphorus in both has shown a little quantity in the ${\gamma}$-ray irradiation in the early period of incubation. After the Ki A embyro, it has increased rapidly and it has increased till the hatching more continually than in control. The about results of the research will be helpful and instructive to the betterment and improvement, breeding and management of animals and plants.

• Journal of Sericultural and Entomological Science
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• v.24 no.2
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• pp.55-72
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• 1983

These studies were done to find out any difference, ultrastructural, physical or chemical, between the shells of diapausing and non-diapausing eggs of the silkworm, Bombyx mori L. 1. From the electron-microscopic observation, the egg shells have four distinctive layers. In addition to the four layers, the shells in the diapausing eggs has another layer with low electron density on its surface. 2. The permeability of the egg shell to hydrochloride was much lower in diapausing egg than in non-diapausing egg. Also the permeability changed in the opposite directions with the egg age: the diapausing eggs decreased while non-diapausing ones increased. 3. The permeability increased when the diapausing egg shell was treated with HCl. When they were treated with ether, however, the increase in permeability was much smaller. It seems there was an ether soluble material involved in the content of the egg shell. 4. The diapausing eggs were also much more resistant to desiccation than the non-diapausing ones. The former, when treated with HCl or chilling, became less resistant to desiccation. 5. The positive histochemical response of the egg shell to PAS-Alcian blue and protein stainings suggests presence of abundant proteins and carbohydrates in the egg shell. On the other hand, the staining response to lipid was more positive in the inner layers than in the outer layer of the shell. 6. The egg shell adhesives seems to be mucopolysaccharides produced by colleterial glands, since the oviposited eggs showed a positive responses to carbohydrate and negative to lipid-staining chemicals, but not the mature oocytes in the ovarioles. 7. There were two bands on the electrophoretic pattern of the SH proteins extracted from the egg shells both in the diapausing egg and non-diapausing one: a slow moving major component and a fast moving minor one. However, the electrophoretic mobility showed a difference in the minor components between them. It is evident that the fast moving minor one of non-diapausing egg ran a little further than that of diapausing egg. 8. In amino acids analysis, no significant differences were found in their composition between diapausing and non-diapausing egg and SH proteins contain relatively more glycine and less cystine.

• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.67-83
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• 1983

The new microsporidia S80 isolated from, Bombyx mori L. in Korea showed ovoid in the morphology of the spores and the size were measured $2.9{\pm}0.28{\mu}$ in length and $1.7{\pm}0.29{\mu}$ width. No other microsporidian spore like this has not been so far isolated from Silkworm. The length of the polar filament extruded in hydrogen peroxide ($H_2O_2$) at $30^{\circ}C$ was $26{\mu}$ of a round cytoplasm on the top. The spores were partly stained with Giemsa, Safranin-O and Gram as the same staining properties as Nosema bombycis, Microsporidia K 79 and other microsporidian spores. The fine structures were observed under scanning eleceron microscope through ultrathin sectioning. The spore wall was composed of three layers ; the thin exospore of an electron dense rippled layer, the thick electron lucent endospore which was thinning considerably at the polar filament insertion point, and the inner limiting membrane. Polar cap present at the sporeapex, with a long polar filament of 12-13 coils, subtending angle of $60^{\circ}$ to spore axis, which is tubular made up of a multilayered and are a benes core, light ring structure enclosing the dance core, the dark ring structure enclosing the inner light ring structure and the other than and light ring structure bounded from cytoplasm. Lamellate polaroplast occupied the anterior part of the spore, and the two neclei with dense nucleoplasm bounded by a double nuclear envelope were cited in the slight downer middle portion of spore. From the characteristics of the shape, size and fine structures, it is certain to reason the Microsporidia S80 belong to the phylum Microspora, class Microspora, order Microsporida, order Microsporida. The shape of two nuclei cited seems to be genus Nosema, but in the classification for the suborder it should be defined wheather pansporoblasts be formed or not and for the genis especial attempts have been made to define the characters which distinguish the disporous genera in the life cycle. Survey through the infection of the bad cocoons during 1980 to 1982 in South Korea the areas contaminated with new microsporidia were revealed 5 provinces of Kyung-Gi, Kang-Won, Chung-Nam and Chun-Nam. Pathological effects inoculated per os at second instar larvae of silkworm, the LD 50 was $7.1{\times}10^7/ml$ as lower pathogenecity than that of Nosema bombycis Naegeli of $1.2{\times}10_7/ml$. While on the other hand the inoculation of the microsporidia at fourth instar larvae lowerd the whole cocoon weight and cocoon shell weight and significant at 1% level. The microsporidia S80 defined it can not be transmitted transovarially from the result of predictive and collective examination of 21 egg batches from the infected female moth.

• Journal of Sericultural and Entomological Science
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• v.21 no.1
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• pp.21-28
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• 1979

The Electrophoretic separation in agarose gel on the esterase and acid phosphatase of blood, midgut and silk gland was carried out with 2 original variginal varieties and 7 F$_1$ hybrids. 1. The midgut of larvae fed on mulberry leaves showed one or two more esterase bands than that of larvae fed on artificial diet. 2. The midgut of C 15 larvae being excellently respondent to artificial diet showed one or two more esterase bands than that of larvae being bad respondent to artificial diet. 3. Electrophoretic separation of esterase bands appeared to be greatly different among newly hatched larvae, 1st and 2nd install larvae of F$_1$hybrids. However the difference among the silkworm varieties was not recognized. 4. According to the change in rearing temperature, the number of the active band of midgut esterase was varied. At the temperature of 28$^{\circ}C$ 5 active esterase bands were found. At temperature of 35$^{\circ}C$ 4 bands were noted at 3rd install and 6 or 7 bands at 4th instar. 5. No similar esterase bands conld be found among midgut, blood and silkgland. There are five esterase bands in the midgut, one in blood and three in silkgland. 6. There was rather small difference in acid phosphatase types of midgut and blood according to varieties and rearing temperature. No active band was shown in silkgland. In midgut, there was one acid phosphatase band at 3rd install, two at 4th instar and three at 5th instar. In blood, One active band at 3rd or 4th instar and three bands at 5th inatar were detected.

• Journal of Sericultural and Entomological Science
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• v.25 no.2
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• pp.1-31
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• 1984

Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.