• 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.

• Parasites, Hosts and Diseases
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• v.48 no.4
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• pp.297-301
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• 2010

Recently, emerging waterbome protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from $10^1$ to $10^2$ oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium paNum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

• Journal of Sericultural and Entomological Science
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• v.28 no.1
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• pp.54-60
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• 1986

Silkworms have been found cross infected with other microsporidia of insects in mulberry trees, forest and fruit gardens. Even the unidentified microsporidian species were not seriously pathogenic to silkworms, the silkworm egg producers lose their profit because of the elimination of eggs laid from moths which are infected with any kind of microsporidian species. Recently, the microsporidian cross infection to silkworm is in tendency of increase and the authors have investigated the field insects to examine the microsporidia. The number of species of insects infected with microsporidia was 10 and they were Boettcherisca peregrina (Robineau-Dewvoidy), Apis melifera linnaeus, Artogenia rapae Linnaeus, Tipula aino Alexander, Altica cacrulescens (Baly), Anomela daimiana Harold, Eilema griseola (Jubner), Rbalbistylun speciosum Uller, Anisodactylus signatus illiger, Oulema oryzae (Kuwayama). From the Boettcherisca peregrina (Rogineau-Desvoidy), three different species of micrsporidia were isolated and the microsporidia isolated from Boettcherisca peregrina (Robineau-Desvoidy), Apis melifera Linnaeus, Artogenia rapae Linnaeus donot have infectivity to silkworm larvae, Bombyx mori L.

• 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.

• 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.

• 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.

• 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.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.87-93
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• 2006

Multiplication and spore production of three microsporidia viz., Nosema bombycis, Nosema sp. 1 and Nosema sp. 2 in fifth instar larval tissues of silkworm, Bombyx mori L. in two seasons with distinct temperature regimes were studied. Nosema sp. 2 produced significantly (P < 0.01) higher number of spores in various tissues. Among the tissues, spore production was highest in silk gland, followed by fat body and gut. Spore production was significantly (P < 0.01) higher in season-II (Average temperature $29.4{\pm}1.1^{\circ}C$). Maximum spore production was observed 25 days post inoculation (p. i.) in season-I (Average temperature $18.9{\pm}1.1^{\circ}C$), whereas in season-II, it was 14 days p. i. In season-I, spore production was low up to 21 days p. i., then increased sharply. In season-II, there was a steady increase in spore production. The results indicate that the microsporidian multiplication is tissue specific and extremely sensitive to temperature at which the host is reared. It also reveals that, silk gland, fat body and gut are the most appropriate tissues for microscopic identification of microsporidia in the larval stage.

• Journal of Sericultural and Entomological Science
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• v.31 no.2
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• pp.113-120
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• 1989

A new microsporidia (S85) was isolated from the silkworm, Bombyx mori L. in Suwon, Korea in 1985. The electron microscopic observation of the fine structure of the spore was studied for the characterization of the new microsporidia and its taxonomic position. The sporewall was composed of three different electron dense layers and the spore surface was rouph and wrinkled. The polaroplast has two parts which differed in the arrangement of the lamellae. The polarfilament was isofilar with thirteen coils and it was consisted of eight concentric layers of different electron density. The spore contained a single nucleus and the pansporoblast appeared as a thin ballon-like structure surrounding the spore. The membrane was generally smooth and shown occationally wrinkled.

• International Journal of Industrial Entomology and Biomaterials
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• v.27 no.2
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• pp.219-227
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• 2013

The purpose of this study was to quickly and effectively identify the Nosema disease of bumblebees in Gangwon Province in Korea. Bumblebees are crucial pollinators of various crops, and microsporidia are the critical pathogens of these hosts. When bumblebees are infected with Nosema bombi, their abdomens can become distended. Paralyzed and infected workers often become sluggish and die early. We have identified the morphology of the microsporidium by light and electron microscopy, and found it to have fairly small oval spores, as has been described previously in many other articles. For the specific and sensitive diagnosis of the microsporidian parasite N. bombi in bumblebees, we have developed an improved method of the polymerase chain reaction (PCR) for expeditious diagnosis. Two pairs of primers were tested on N. bombi and the related microsporidia Nosema apis and Nosema sp., both of which infect Bombus ignitus and Bombus hypocrita sapporoensis. Furthermore, we have verified and analyzed the 16SrRNA sequence data of N. bombi in bumblebees by using the Basic Local Alignment Search Tool (BLAST) server at the National Center for Biotechnology Information.