• Journal of Sericultural and Entomological Science
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• v.39 no.2
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• pp.146-153
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• 1997

The microsporidian infection with Nosema bombycis, reconfirmed its high virulence and transovarial tranmissibility, however, the characteristic symptom of the spots like pepper grains on the diseased larval skin was no more recognized by present varieties of the silkworm. Transovarial transmission rate detected from moth was above 90% in dead eggs or dead larvae in the rearing by mulberry leaves, 80% in the newly hatched larvae starved to death. Transovarially transmitted N. bombycis was easily observed from dead eggs and larvae, and were suggested an individual inspection of a few of dead eggs for detection of the pathogenic spores. The progeny population provided indicative factors on the sampling of predictive and corrective inspection. The higher concentration of N. bombycis spores included in the hindabdormal part of infected moth, applicative on the simple method of indivisual moth inspection. For the predictive inspection of growned 5th-instar larvae, N. bombycis infection was detectable without microscopic observation by the unique symptom of turbid milky-white spots on the silk gland. Inspection of the meconia artificially discharged from silkworm moth, was also succesful of microscopic observation before crossing, without killing or homogenize the moths. The results provided a basis of rational methods for the inspection of N. bombycis infection of the silkworm.

• Journal of Sericultural and Entomological Science
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• v.39 no.2
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• pp.154-160
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• 1997

Present experiment designed for the review of theoretical basis for the inspection system of infected insects by Nosema bombycis. A microporidian N. bombycis, known as the high virulence, produced at the average of 7$\times$108 spores per female moth of the silkworm, Bombyx mori, enabled transovarial tranmission. Detectability of N. bombycis spores in the mass inspection was varied by dillution level, the higher limit of dillution with healthy moths was 1:140 for 100% detection, 1:160 for 99.5%, 1:200 for 99.0%. For an efficient inspection under the microscopic observation (600$\times$), the lower limit of spore concentration was determined as 1, 000, 000 spores/ml, 60-80 moths could be applicable for a maximum sample unit of a lot. Following the present inspection unit conditioned 35 to 40 moths for a lot. N. bombycis spores were easily detectable from the preparation of crude homogenate with 2% KOH, even the step of contrifuge was omitted. The results suggested a new basis of rational mass inspection system of silkworm female moths to save the facilities, labor, and time.

• Journal of Sericultural and Entomological Science
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• v.38 no.1
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• pp.48-52
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• 1996

Nosema bombycis was found on the both of surface and inside of silkworm eggs layed by infected female moth, maternal-mediated transmission of those pathogens via the surface or internal site of silkworm eggs were investigated. All of the meconia from infected female moths contained pathogenic spores, those concentration were 4.6(${\pm}$0.24)X106 /ml. The pathogens on the surface of nondiapause eggs were transmitted to the progeny larvae at the rate of 54.5%, however, lost their activity before hatching for the case of overwintered eggs or acid treatment(16% HCI at 46.2$^{\circ}C$ for 6 min). N. bombycis in the silkworm eggs localized at the serosa, yolk, embryo except chorion tissue in the silkworm eggs layed by infected female moths. Development of the pathognes in the eggs was synchronized with embryogenesis, which secured the safety of pathogens against environmental condition, resulting in the high transmissibility of 91.2${\pm}$1.80%.

• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.2
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• pp.107-111
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• 2005

Impact of microsporidian infection and season on reproductive potential of Bombyx mori L. was investigated in the laboratory. Microsporidian infection significantly (P<0.01) reduced fecundity and hatching and increased sterility and mortality of eggs. Among the microsporidia, Nosema sp. 2 infected silkworm produced eggs with least fecundity and hatching percent as well as highest dead and sterile eggs followed by Nosema sp. 1 and N. bombycis. Microsporidia, in general, significantly reduced fecundity and hatching percent of eggs and increased number of dead and sterile eggs in all the three seasons except N. bombycis in July - August (S3) and Nosema sp. 2 in January February (S1). Since, seed production is the anchor sheet of mulberry sericulture, coefficient of egg lying is considered as an important aspect and the industry quite often facing shortage of disease free layings. The present study indicates that B. mori is more susceptible to microsporidia during S2 followed by S3 and S1 and Nosema sp2. is most virulent followed by Nosema spl. and N. bombycis.

• Journal of Sericultural and Entomological Science
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• v.36 no.1
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• pp.69-75
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• 1994

Through the histological and anatomical investigation of silkworm gonad, N. bombycis infection was found to begin from the peripheral region of ovarial sheath or testicular sheath, then, the pathogens spread to the inner portion. Peroral inoculation with purified spores of N. bombycis to 2nd instar larvae at dosages around 106-8/㎤ of artificial diet resulted in the extremely extended larval survival as long as 15 to 22 days of 4th instar. The growth of ovarioles was confirmed in the 10 to 14 day old larvae, oogonia developed into oocytes and nurse cell against heavy infection of the ovary. Gonads rarely obstructed oogenesis and spermatogenesis in the pupae failed in adult eclosion. Light infection of female hosts effected insignificantly on the ovarial development, however, recorded 100% transmission of the pathogens to the progeny populations. Conclusively, ovarial inflection of silkworm induce transovarial transmission begins around 2~3 day old pupae when ovarioles extruded out to hemocoel, and the infection period thought to be continued until the stage of eggs complete shell formation in the ovariole.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.2
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• pp.83-89
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• 2009

Multiplication and spore production of three microsporidia(Nosema bombycis, Nosema sp. 1 and Nosema sp. 2) in selected tissues of pupa and adult of silkworm, Bombyx mori L. were studied in two seasons (SI, SII) with distinct temperature (SI: $20.1{\pm}0.8^{\circ}C$ and SII: $25.1{\pm}0.7^{\circ}C$) regimes. Multiplication of the microsporidia followed a logistic pattern with a lag phase, an exponential phase and a stationary phase. In SII, spore production was significantly (P<0.01) higher in various tissues. Highest spore production was observed 30 days post inoculation (p.i.) in SI and in SII, it was $21{\sim}23$ days p.i. Spore production was significantly (P<0.01) higher in the gut tissues than other tissues. Nosema sp. 2 registered significantly (P<0.01) higher spore production in both the seasons compared to Nosema bombycis and Nosema sp. 1. Results indicate that the multiplication and spore production of microsporidia are tissue specific and extremely sensitive to the temperature at which the host is reared. Through this study, the precise day that the spore numbers of the microsporidia are maximized can be predicted in both pupa and adult in case the infection is initiated in the first instar.

• International Journal of Industrial Entomology and Biomaterials
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• v.17 no.1
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• pp.157-163
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• 2008

Infection of pathogenic microsporidia, Nosema bombycis and Nosema mylitta (Chakrabarti and Manna, 2006) decreased egg production, fecundity, hatching % and increased sterile eggs in heavily infected mulberry silkmoth, Bombyx mori L. On an average a disease free moth laid upto 442.67 eggs with high hatching % (99.53) and less sterile eggs ($0.47{\sim}2.00%$). While an infected moth laid less number of eggs ($7.00{\sim}412.00$) with low hatching % ($32.437{\sim}98.643$) and high sterile eggs ($2.143{\sim}129.571$). Fecundity of disease free laying was highest (468.714) during season-1 then gradually decreased during season- 2 (414.000) to season- 3 (404.285). But fecundity of an infected laying was highest during season-2 and hatched eggs were lowest during season-2. Higher inoculums concentration of N. mylitta infected to 5th stage larva of mulberry silkworm drastically decreased the fecundity in season - 3 and lower inoculums concentration of N. bombycis decreased the fecundity in season-1 and 3. Season-3 was most effective season to decrease the fecundity and increase sterile eggs when both temperature and humidity were fluctuated from the optimum level.

• International Journal of Industrial Entomology and Biomaterials
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• v.27 no.2
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• pp.282-288
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• 2013

In nature, the population of Nosema bombycis (Microsporidia) causing pebrine disease is small and their development is extremely slow and only few ultimately producing spores. Pebrine infected silkworm, Bombyx mori larvae collected from sericulture field were alive till $3^{rd}$ generation though the concentration of N.bombycis spore was very high ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$). All larvae were died during $4^{th}$ generation with extremely high concentration of pebrine spores ($3.0-4.0{\times}10^9$ spores. $mL^{-1}$) and mostly contain long polar tube (LT). Alternately, all larvae were died immediately (at $3^{rd}$ stage of $1^{st}$ generation) when it was artificially inoculated with same concentration of N.bombycis spores harvested from field ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$) though concentration of spores harvest was very less ($3.0-4.0{\times}10^6$ spores. $mL^{-1}$) and mostly contain short polar tube (ST). Artificially pebrine infected male moth when mated with healthy female moth took six generations to develop pebrine disease and all larvae were died at the $2^{nd}$ stage with very less spore harvest ($3.0-10.0{\times}10^6$ spores. $mL^{-1}$). Survival percentage was increased in all generations (~92.0% at $4^{th}$ generation) when silkworm rearing was conducted under new integrated disease management system.

• Journal of Sericultural and Entomological Science
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• v.27 no.1
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• pp.1-11
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• 1985

In order to clarify the taxanomic position of a new microsporidia K79 which was isolated from the silkworm larvae, Bombyx mori L. in Korea in 1979, the following several experiments such as estimation of pathogenicity in different instar, histopathological examination under light and electronic microscope and examination of fine structure of the sporse were carried out and their result obtained are as follows. In the test of pathogenicity by oral inoculation, the new microsporidia K79 was lower than Nosema bombycis and the susceptibility of the new microsporidia to silkworm was getting lower as the silkworm larvae grew. The lesion of Silkworms' tissue which was infected with the new microsporidia K79 was found in the epithelial cells of trachea, fat body and silk gland cells. The developmental process of the new microsporidia K79 in vivo could be divided into the following five stages: sporoplasm, schizont, sporont, sporoblast, and spore. The process was just the same as the of N. bombycis, but its development was slower than that of N. bombycis. Several differences in the fine structure of the spore under electron microscope, which could be important keys for the classification of microsporidia, were obtained. Anchoring disk and polaroplast lamella of the new microsporidian spore were disclosed to be different from those of N. bombycis. An average number of polar filament coils of the new microporidian spore was 16 at an angle of 75$^{\circ}$. On the basis of various keys for the classification of microsporidia, the results obtained from various experiments proved that the newly isolated microsporidia should be classified into the Genus, "Nosema", nut is further classification for species should be conducted in the future. Therefore, it may be reasonable that the new microsporidia is temperally classified as Nosema sp. K79 considering the fact that it was discovered in Korea in 1979.a in 1979.

• 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.