• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.737-744
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• 2004

A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Vairimorpha spp. and Nosema spp. and identification of Vairimorpha necatrix from Lepidoptera insects. Three sets of primers were selected from different genomic sequences to specifically amplify an 831 bp amplicon within the SSU rRNA gene, specific for both Vairimorpha spp. and Nosema spp. (MSSR primer); a 542 bp amplicon within the SSU rRNA gene, specific for Vairimorpha spp. (VSSU primer); and a 476 bp amplicon within the actin gene, specific for Vairimorpha necatrix (VNAG primer). Using the primers in conjunction with multiplex PCR, it was possible to detect Vairimorpha spp. and Nosema spp. and to differentiate between them. The sensitivity of this PCR assay was approximately 10 spores per milliliter. It is proposed that the multiplex PCR is a sensitive, specific, and rapid tool that can serve as a useful differential diagnostic tool for detecting Vairimorpha spp. and Nosema spp. in Lepidoptera insect.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2001년도 추계학술대회
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• pp.77-81
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• 2001

Enterocytozoon bieneusi는 특히 AIDS환자에게 치명적인 질병을 초래하는 세균과 원충 중간단계의 병원체로서 현재 $30\~50\%$의 발생률오 AIDS환자로부터 가장 많이 발생되는 병원체로 알려져 있다. 하지만 이 병원체는 1980년 후반에서야 비로서 학계의 주목을 받기 시작했으며 환자에게 심한 만성설사, 기력 쇠진, 악성 영양 결핍 등을 주 임상증상으로 하고 있다. 이렇게 다양하고 심각한 질병을 일으킴에도 불구하고 현재까지 이 병원체에 대한 연구가 미흡하였다. 현재 이 병원체에 대한 가장 중요한 연구 중 연구 중 하나는 이 병원체에 대한 AIDS환자로의 전파 경로이다. 최근 여러 각도의 동물야외 조사 및 실험동물을 이용한 연구를 토대로 AIDS환자에 대한 이 병원체의 전파가능성은 감염동물에서부터 전파가 가장 중요한 경로가 될 거라 보고되고 있다.

• 한국잠사곤충학회지
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• 제38권1호
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• pp.48-52
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• 1996

Nosema bombycis의 경란전염은 집누에의 경우 난내 및 난면의 두가지 경로가 존재하는 것으로 입증되었다. 난내에서 병원체가 존재하는 부위가 난각 조직을 제외한 장액막, 난황, 배자 등이였다. 난내에 침입한 병원체는 난의 발육과 동조적으로 발육 증식하므로써 오랜 시일이 경과하여도 불활화 되지 않는 동시에 감염난의 치사를 줄일 수 있으며, 병원체가 안전하고 확실하게 차대의 유충으로 전달되므로난내경로는 경난전염율이 높은 것으로 밝혀졌다. 난면경로는 감염 모체의 아충에 의한 난 표면의 오염이 주 원인으로서, N. bombycis 감염아는 100%가 아뇨에 병원체가 검출되며, 4.6$\pm$0.24)X106/ml의 포자를 함유하는 것으로 밝혀졌다. 난 표면에 부착된 병원포자는 비휴면란에 도말하였을 때, 10일 정도 경과 후 부화된 유충의 54.5%가 감염되었으나, 월년 보호하거나 침산한 경우 차대잠의 감염은 인정되지 않았다. 따라서, 집누에 월년란의 경우 경란전염에 의한 질병의 만연에는 난내경로가 주요한 것으로 결론지었다.

• 한국잠사곤충학회지
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• 제36권1호
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• pp.69-75
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• 1994

경란전염은 숙주의 생식소 감염이 전제되므로 Nosema bombycis 감염 집누에를 대상으로 생식소의 발육에 수반된 란 및 정자 형성과 경란전염의 성립에 미치는 영향을 해부 및 조직병리학적 측면에서 구명하였다. 생식소의 감염경로는 도관을 통하지않고 체공에 접한 주변부의 피막조직이 먼저 감염된후 난소내부로 병세가 확산되었고, 감염시기는 다른 조직에 비하여 지연되므로 중증인 개체에 한하여 생식소의 감염은 인정되었으나 란 및 정자 형성이 저해되고 숙주는 성충화 전에 치사하였다. 생식능을 보유한 경증의 감염개체에서는 난소의 감염이 번데기 초기까지 인정되지 않았으나 경란전달은 가능하였다. 병원포자는 자력으로 이동할 수 없으며 성숙란에 대한 침입 가능성도 부정적인 것으로 판단되었으므로, 경란전염이 성립되는 란의 감염시기는 난소소관이 체공에 직접 노출되는 번데기 시기의 2~3일경 부터 난소난의 난각형성이 개시되기 전인 제10 발육단계까지의 기간이 될 것으로 추정되었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제29권2호
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• pp.169-178
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• 2014

In this study, we investigated genetic diversity of 22 microsporidian isolates infecting tropical tasar silkworm, Antheraea mylitta collected from various geographical forest locations in the state of Jharkhand, India, using polymerase chain reaction (PCR)-based marker assay: random amplified polymorphic DNA (RAPD). A type species, NIK-1s_mys was used as control for comparison. The shape of mature microsporidians was found to be oval to elongate, measuring 3.80 to $5.10{\mu}m$ in length and 2.56 to $3.30{\mu}m$ in width. Of the 20 RAPD primers screened, 16 primers generated reproducible profiles with 298 polymorphic fragments displaying high degree of polymorphism (97%). A total of 14 RAPD primers produced 45 unique putative genetic markers, which were used to differentiate the microsporidians. Calculation of genetic distance coefficients based on dice coefficient method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was conducted to unravel the genetic diversity of microsporidians infecting tasar silkworm. The similarity coefficients varied from 0.059 to 0.980. UPGMA analysis generated a dendrogram with four microsporidian groups, which appear to be different from each other as well as from NIK-1s_mys. Two-dimensional distribution based on Euclidean distance matrix also revealed considerable variability among different microsporidians identified from the tasar silkworms. Clustering of few microsporidian isolates was in accordance with the geographic origin. The results indicate that the RAPD profiles and specific/unique genetic markers can be used for differentiating as well as to identify different microsporidians with considerable accuracy.

• International Journal of Industrial Entomology and Biomaterials
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• 제27권2호
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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.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
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• pp.6-6
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• 2023

Nosemosis is one of the most common protozoan diseases of adult bees (Apis mellifera). Nosemosis is caused by two species of microsporidia; Nosema apis and Nosema ceranae. Nosema ceranae is potentially more dangerous because it has the ability to infect multiple cell types, and it is now the predominant microsporidian species in A. mellifera. In this study, we identified two anti-nosemosis plants, Aster scaber and Artemisia dubia, which reduced the spore development of N. ceranae in spore-infected cells. We intend to establish the anti-nosemosis activity of aqueous, ethyl acetate (EA), and butanol (BuOH) extracts of A. dubia and A. scaber. In order to determine the optimal dose, we did in vitro and in vivo toxicity for all the extracts and carried out anti-nosemosis experiments. Although all of the extracts (aqueous, EA, and BuOH) showed in vitro and in vivo anti-nosemosis activity in a dose-dependent manner, the aqueous extracts of A. dubia and A. scaber showed more potent anti-nosemosis activity than the EA and BuOH extracts. And then, we isolated five phenolic compounds [chlorogenic acid, 3,4-dicaffaeoylquinic acid (3,4-DCQA), 3,5-dicaffaeoylquinic acid (3,5-DCQA), 4,5-dicaffaeoylquinic acid (4,5-DCQA), and coumarin] from A. dubia, A. scaber, and A. dubia + A. scaber aqueous extracts and screened for their toxicities and anti-Nosema effects in both in vivo and in vitro conditions. Among these five compounds, coumarin, chlorogenic acid, and 4,5-DCQA exhibited less toxic but more potent anti-Nosema effects than the other two compounds. Especially, chlorogenic acid and coumarin showed prominent anti-Nosema activities even at the lowest concentration (10 ㎍/mL). They might have potential to be developed as alternative compounds for the control of Nosema disease.

• Journal of Microbiology and Biotechnology
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• 제34권5호
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• pp.1146-1153
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• 2024

The increasing economic losses associated with growth retardation caused by Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp, require effective monitoring. The internal transcribed spacer (ITS)-1 region, the non-coding region of ribosomal clusters between 18S and 5.8S rRNA genes, is widely used in phylogenetic studies due to its high variability. In this study, the ITS-1 region sequence (~600-bp) of EHP was first identified, and primers for a polymerase chain reaction (PCR) assay targeting that sequence were designed. A newly developed nested-PCR method successfully detected the EHP in various shrimp (Penaeus vannamei and P. monodon) and related samples, including water and feces collected from Indonesia, Thailand, South Korea, India, and Malaysia. The primers did not cross-react with other hosts and pathogens, and this PCR assay is more sensitive than existing PCR detection methods targeting the small subunit ribosomal RNA (SSU rRNA) and spore wall protein (SWP) genes. Phylogenetic analysis based on the ITS-1 sequences indicated that the Indonesian strain was distinct (86.2% nucleotide sequence identity) from other strains collected from Thailand and South Korea, and also showed the internal diversity among Thailand (N = 7, divided into four branches) and South Korean (N = 5, divided into two branches) samples. The results revealed the ability of the ITS-1 region to determine the genetic diversity of EHP from different geographical origins.