농작물의 수분 매개자이며 생태계 유지에 필수적인 역할을 하는 꿀벌의 노제마병(nosemosis)은 꿀벌 집단 붕괴현상(colony collapse disorder: CCD)의 원인 중 하나로 알려져 있다. 또한, 노제마병은 봉군 약화를 초래할 뿐 아니라 여러 가지 양봉 산물의 생산성을 낮추는 원인이기도 하다. 국내에서는 Nosema ceranae가 노제마병의 주요 병원체로 알려져 있다. 노제마 감염을 확인하기 위해서는 꿀벌의 중장을 적출한 후 노제마 포자의 확인 및 계수를 통해 감염 수준을 평가할 수 있다. 본 연구에서는 더욱 빠르고 정확하게 노제마 감염 수준을 평가할 수 있는 새로운 방법으로 포자 염색법과 qPCR 방법을 개발하였다. 노제마 포자에 대해 특이적인 염색이 가능한 Fluorescent Brightener를 이용하여 노제마 감염 꿀벌 중장을 염색한 결과, 형광 발현으로 노제마의 감염 여부를 확인할 수 있었다. 또한, 중장내 포자량에 비례한 형광 발현도 확인할 수 있었다. 그러나, 노제마 감염에 대한 특이도 및 포자량에 비례한 형광 발현 민감도는 신뢰하기 어려웠다. 그에 비해, 노제마 특이 유전자를 이용한 qPCR 방법은 노제마의 감염 여부 뿐만 아니라 포자량에 비례한 감염 수준 결정이 가능함을 확인하였다. 특히, 많은 시료들에서 노제마 감염 수준의 신속하고 정확한 평가에 qPCR 방법은 유용하게 활용될 수 있을 것으로 기대되었다.

Nosema disease caused by the microsporidia Nosema apis and Nosema ceranae are a honey bee pathogen parasitizing. Nosema disease symptoms include digestive and absorption disorders because the spores damage epithelial tissue and potentially causing colony death. Recently, N. ceranae has been reported as an important threat to honey bee health. Turmeric (Curcuma longa L.) Curcuma tonga L. belongs to the family Zingiberaceae and is a perennial, tropical herb. Turmeric, the powdered rhizome, is used for medicinal purposes. The aim of this study was to evaluate the potential of Turmeric (Curcuma longa L.) for the control of N. ceranae in honeybees. For the study, we infected with N. ceranae spore through dosed and fed with the turmeric extraction at difference concentration. The data show that the turmeric extraction was not toxic for bee at least at 1% and the bees fed with 0.5 % turmeric extraction had significantly lower infection rates. This data suggest that turmeric could be useful in alternative strategies for the control of N. ceranae.

The population of managed honeybees has been dramatically declining the recent past in worldwide. N. ceranae causessignificant detriment to honey production and results in economic losses critically. In our knowledge, Fumagillin is theonly antibiotic approved for control of nosemosis in honeybees. In this study, to select isolate with anti-Nosema activityagainst N. ceranae. Entomopathogenic fungi cultural filtrates were screened using in vitro polar tube germination assay.These fungi cultural filtrates were used to evaluate the safety of honeybees and their inhibition of nosemosis. As a result,P. marquandii 364 and Pochonia sp. 60 showed inhibitory activity on the growth of N. ceranae in honeybees and didnot significantly affect the survival rate of honeybees. These may be employed as antibiotic agents and a good featureto be used in the development of new biocontrol agents of nosemosis.

Honeybees have faced many diseases which treaten bee colony including a serious population decline phenomenon called Colony Collapse Disorder (CCD). Nosema ceranae is a pathogen cause nosemosis, which is now wide spread around the world. According to the genome sequencing for N. ceranae, it has been identified that the presence of machinery for RNA silencing. Microsporidia N. ceranae that are obligated intracellular parasites depend on their host for energetic and metabolic needs. Here we selected the several genes from mitosome of N. ceranae to develop RNAi for the control of Nosema. Especially, TOM40, FNR1, FNR2 and Nar1 were chosen. After infection of N. ceranae, the Honeybee were treated with RNAi either by using only one or combining two or more. The infection rate and specific gene silecing in Nosema were analyzed.

The bumblebee, Bombus terrestris, has played an important role as one of the alternative pollinators since the outbreak of honeybee collapse disorder. Recently, pathogens and parasites such as viruses, bacteria and mites, which affect the life span and fecundity of their host, have been discovered in B. terristris. In order to detect the microsporidian pathogen, Nosema Spp. in the field populations of B. terristris, we collected adults and isolated their genomic DNA for diagnostic PCR. The PCR primers specific for Nosema Spp. were newly designed and applied to gene amplification for cloning. Only small subunit ribosomal RNA(SSU rRNA) gene of N. ceranae was successfully amplified and sequenced among examined genes, which indicates that N. ceranae mainly infects the examined field population of B. terristris. To detect of SSU rRNA gene, two regions of SSU rRNA gene were selected by primary PCR analysis and further analyzed in quantitative real-time PCR(qRT-PCR). The qRT-PCR analysis demonstrated that SSU rRNA of N. ceranae was detected at concentrations as low as 0.85 ng/μl genomic DNA. This result suggests that the detection via qRT-PCR can be applied for the rapid and sensitive diagnosis of N. ceranae infection in the field population as well as risk assessment of B. terristris.

A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Nosema apis and Nosema ceranae in honeybee. Three sets of primers were selected from different genomic sequences to specifically amplify a 831 bp amplicon within the SSU rRNA gene, specific for both N. apis and N. ceranae (MSSR primer); a 375 bp amplicon within the SSU rRNA gene, specific for N. apis (NA primer); and a 1,131 bp amplicon within SSU rRNA gene, specific for N. ceranae (NC primer). Using the primers in conjunction (multiplex PCR) we were able to N. apis and N. ceranae and to differentiate between them. The sensitivity of this PCR assay was approximately 102 spores per milliliter. We proposed that the multiplex PCR was sensitive, specific and rapid tool that can serve as a useful differential diagnostic tool for detecting N. apis and N. ceranae in honeybee.

A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Nosema apis and Nosema ceranae in honeybee. Three sets of primers were selected from different genomic sequences to specifically amplify a 831 bp amplicon within the SSU rRNA gene, specific for both N. apis and N. ceranae (MSSR primer); a 375 bp amplicon within the SSU rRNA gene, specific for N. apis (NA primer); and a 1,131 bp amplicon with in SSU rRNA gene, specific for N. ceranae (NC primer). Using the primers in conjunction (multiplex PCR) we were able to N. apis and N. ceranae and to differentiate between them. The sensitivity of this PCR assay was approximately 102spores per milliliter. We proposed that the multiplex PCR was sensitive, specific and rapid tool that can serve as a useful differential diagnostic tool for detecting N. apis and N. ceranae in honeybee.