• Parasites, Hosts and Diseases
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• 제59권6호
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• pp.639-643
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• 2021

Enterocytozoon bieneusi is a microsporidian pathogen. Recently, the equestrian population is increasing in Korea. The horse-related zoonotic pathogens, including E. bieneusi, are concerns of public health. A total of 1,200 horse fecal samples were collected from riding centers and breeding farms in Jeju Island and inland areas. Of the fecal samples 15 (1.3%) were PCR positive for E. bieneusi. Interestingly, all positive samples came from Jeju Island. Diarrhea and infection in foals were related. Two genotypes (horse1, horse2) were identified as possible zoonotic groups requiring continuous monitoring.

• 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.

• 한국잠사곤충학회지
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• 제39권2호
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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.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권1호
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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.

• Parasites, Hosts and Diseases
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• 제52권3호
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• pp.321-323
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• 2014

Encephalitozoon cuniculi is a microsporidian parasite commonly found in rabbits that can infect humans, causing encephalitozoonosis. The prevalence of encephalitozoonosis is not well documented, even when many clinics suspect pet rabbits as being highly infected. This study investigated the seropositivity of E. cuniculi using ELISA. The examination of 186 rabbits using ELISA showed that 22.6% (42/186) were seropositive against E. cuniculi. In analysis with healthy status, all 42 seropositive sera were collected from clinically normal rabbits. Moreover, the gender and age of pet rabbits did not have anysignificant effect on E. cuniculi infection. To the best of our knowledge, this is the first report to describe the seroprevalence of E. cuniculi in pet rabbits and suggests that pet rabbits could act as an important reservoir of encephalitozoonosis for both pet animals and humans in Korea.

• 한국양봉학회지
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• 제34권2호
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• pp.137-140
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• 2019

Although honeybees (Apis mellifera) are crucial for maintenance of the ecosystem, population of honeybee has been steadily decreasing due to diseases including nosemosis. Nosemosis is a disease caused by Nosema ceranae and is now considered as a major threat to honeybees. N. ceranae is a microsporidian that stays in form of spore even before the infection, which makes it harder to control than other pathogens. People are now aware of this parasite, however, cure and preventive candidates for nosemosis are hardly found until today. In this study, in vitro experiment of Lespedeza cuneata treatment to prevent nosemosis were done using Trichoplusia ni cell line, BTI-TN5B1-4. Normal T. ni cells exhibited round shape without abnormal size. On the other hand, when N. ceranae were treated, cells deteriorated and some cells abnormally enlarged due to N. ceranae infection. Interestingly, treatment of T. ni cells with L. cuneate extract protected abnormal cell shape induced by N. ceranae infection to normal shape. Some N. ceranae spores were observed outside of the cells. Effective concentration range for N. ceranae control were experimented. Lowest concentration which can control nosemosis were 50 ㎍/mL. When the concentration of L. cuneata extract was exceeded 200 ㎍/mL, cytotoxicity started to show up.

• 한국임상수의학회지
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• 제32권2호
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• pp.212-214
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• 2015

토끼에서 뇌포자육충증은 Encephalitozoon(E.) cuniculi에 의해 유발되는 전염성 질환이다. E.cuniculi는 숙주영역이 상당히 넓으며, 특히 면역능이 저하된 사람에서 기회 감염을 유발하기도 한다. 본 증례 보고에서 4 마리의 토끼가 사경과 안구진탕 증상으로 내원하였으며, 각 개체의 채혈을 통해 ELISA 검사를 수행하여 E.cuniculi에 양성임을 확인하였다. 토끼들은 fenbendazole로 4 주간 치료를 실시하였고, 치료효과는 신경증상의 발현정도에 따라 평가하였다. 치료한 네 마리 토끼 중 세 마리의 토끼에서 치료 3-5일 후부터 증상이 상당히 개선되었으나, 한 마리는 식욕부진과 사경증상이 개선되지 않고 치료 후 10일째 폐사하였다. 본 증례보고는 뇌포자육충 감염 애완토끼에 대한 fenbendazole의 치료효과를 규명한 최초의 보고이며, 본 증례를 통하여 애완토끼가 사람과 동물에 있어서 잠재적 E. cuniculi 보균동물로 인식되므로 이에 대한 주의를 필요로 한다.

• International Journal of Industrial Entomology and Biomaterials
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• 제27권2호
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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.

• Journal of Veterinary Science
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• 제22권3호
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• pp.40.1-40.12
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• 2021

Background: The microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen. Objectives: The present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees. Methods: A procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration. Results: UR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min. Conclusions: UR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.

• 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.