냉수성 질병에 대한 제주 양식 돌돔, Oplegnathus fasciatus의 병리조직학적 관찰

문경미 ( Kyung Mi Moon ),고대희 ( Dae Hee Ko ),김동휘 ( Dong Hwi Kim ),박근태 ( Geun Tae Park ),허문수 ( Moon Soo Heo ) 한국어병학회 2013 한국어병학회지 Vol.26 No.2

The outbreak of haemorrhagic speticemia and dermal lesions in cultured Rock sea breem in Jeju Island are frequently associated with keratitis. Prolonged infection leads to the symptoms accompanied by necrosis of fin and skin and the fishes died. Present study aims to study the histopathological changes of diseased fish muscle tissue. The tissue at the initial stage of infection showed normal muscular texture, however in the late stages of infection, muscular lesion had been expanded and hence, necrosis had progressed deeply into muscle tissue. Mild tissue in caudal fin was found with more necrosis, and gill tissues were also collapsed. In Kidney, renal tubules were teared along with tissue destruction (Espada, J et al, 1993). Hence in the present study, we examined the histopathological variation of the infection fishes, and this basic data would be used for future research of in haemorrhagic specticemia and dermal diseases.

한국전통식품으로부터 분리 된 세균의 항균활성 효과

문경미(Kyung-Mi Moon),허문수(Moon-Soo Heo) 한국생명과학회 2015 생명과학회지 Vol.25 No.11

매년 어류 양식의 생산량은 증대되고 있으며, 생산량 증가를 위한 고밀도 사육은 수질 악화 및 어류들의 스트레스를 증가시켜 질병들이 잦아지게 된다. 이로 인한 경제적 피해는 어류 양식 어민들에게 있어 막대한 피해를 입게되며, 이를 예방하기 위해 화학 요법인 항생제를 투여하게 된다. 본 연구에서는 항생제 대체 물질 및 안전 요법으로 알려진 프로바이오틱스를 분리하기 위해, 한국전통식품인 고추 장아찌와 갈치 젓갈에서 다양한 세균을 분리하였다. 분리 된 균주들은 16S rDNA 염기서열을 분석 및 동정하고, 상층액 및 균체로 나뉘어 항균 활성을 측정하였다. 고추 장아찌에서 분리된 4종과 갈치 젓갈에서 분리된 7종은 상층액 보다 균체에서 항균 활성이 높게 나타나는 것을 확인할 수 있었다. 그 중 JKM-2는 43 mm(S. iniae), 40 mm(S. parauberis), 35 mm(S. mutans), 26.5 mm(V. vuinificus)로 가장 높은 항균 활성을 나타냈다. 본 균주의 염기서열 분석 결과, Bacillus subtilis 97.71%, Bacillus tequilensis 97.71%, Brevibacterium halotolerans 97.71%, Bacilus subtils 97.63%, Bacillus subtlis 97.63%, Bacillus mojavensis 97.54%, Bacillus vallismortis 97.46%, Bacillus nanillea 97.45%, Bacillu smethylotrophicus 97.37%, Bacillu ssiamensis 97.37%로 분석되었다. 추후 JKM-3의 신종 균주 확인과 균체 물질 규명 및 분석을 통하여 충분한 안정성을 확보하고 양식 산업에 적용시켜 항생제 대체 물질로서의 이용가치를 확인하고자 한다. Aquaculture continues to be an ever-growing sector. However, high-density farming increases disease outbreaks due to deteriorating water quality and internal stress. To prevent disease, the most common method chemotherapy is using antibiotic administration. In this study, probiotic bacteria were isolated from Korean traditional foods, such a Gochu pickle and cutlassfish salted seafood. Various bacteria were isolated, and their 16S rDNA sequences were analyzed. The antimicrobial activities of four isolates from Gochu pickle and seven isolates from cutlassfish salted seafood were assayed, in addition to the antibacterial activity of culture pellet and supernatant. The antibacterial activity of the pellet was higher than that of the supernatant. Isolate JKM-2 showed the highest antibacterial activity against Streptococcus iniae (43 mm), S. parauberis (40 mm), S. mutans (35 mm), and Vibrio vuinificus (26.5 mm). The sequences of the isolated strains were compared with those of Bacillus subtilis (97.71%), B. tequilensis (97.71%), Brevibacterium halotolerans (97.71%), B. subtilis (97.63%), B. subtilis (97.63%), B. mojavensis (97.54%), B. vallismortis (97.46%), B. nanillea (97.45%), B. methylotrophicus (97.37%), and B. ssiamensis (97.37%). Future through analysis and new strains confirmed the bacterial cell material investigation of JKM-3, and to ensure sufficient stability, it is desired to verify the utility value as a substitute material for antibiotics by application to the form of the industry.

Loop-mediated isothermal amplification (LAMP)법을 이용한 Streptococcus parauberis의 신속 진단

문경미(Kyung-Mi Moon),김동휘(Dong-Hwi Kim),허문수(Moon-Soo Heo) 한국생명과학회 2014 생명과학회지 Vol.24 No.4

Loop-mediated isothermal amplification (LAMP)법은 등온에서 DNA 주형을 변성시키지 않고 실시하기 때문에, autocycling 가닥 변위 DNA 합성에 의존한다. 그래서 고가의 PCR 장비를 필요로 하지 않고 등온 유지가 가능한 저가의 장비인 항온 수조, 오븐, 온장고 등에서 증폭이 가능하다. 본 연구진은 Streptococcus parauberis의 random primer중에서 5개를 선정하여, 신장도가 높은 2개의 primer를 이용하여 최적 반응온도 및 최적 반응시간, 최적 반응 조건들을 확립하였다. 그리고 기존의 PCR과 LAMP의 민감도의 비교 분석을 측정한 결과, LAMP의 높은 검출 한계를 확인할 수 있었다. 본 논문에서는 non-target DNA의 영향을 받지 않고 등온 조건 하에서 DNA를 증폭시킬 수 있는 LAMP법과 SYBR-green I를 이용하여 시각화시켰으며, 기존의 PCR과 비교 분석함으로써, S. parauberis에 대한 신속하고 정확한 진단법을 확립하였다. Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR. Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.

제주 연안의 괭생이모자반(Sargassum horneri)에서 분리된 세균의 계통학적 다양성 및 군집 구조 분석

문경미(Kyung-Mi Moon),박소현(So-Hyun Park),허문수(Moon-Soo Heo) 한국생명과학회 2018 생명과학회지 Vol.28 No.10

최근 괭생이모자반(Sargassum horneri)은 제주의 해안가와 해변, 연근해 양식장 등 매년 대량으로 속출되어 인근 양식업자 및 주민에게 막대한 피해를 주고 있는 추세이다. 본 연구에서는 제주 인근 연안으로 흘러 들어온 괭생이모자반에 서식하고 있는 미생물의 다양성을 탐색하고 동정을 통한 미생물의 기능을 파악함으로써 생태학적 문제에 관한 기초자료를 제공하고자 하였다. Proteobacteria는 분리 된 균주 중 88%를 차지한 우점문으로 α-proteobacteria강은 6속 10종으로 Pseudorhodobacter속이 40%를 차지하고 Paracoccus속 20%, Rhizobium, Albirhodobacter, Skermanella 및 Novosphingobium 속은 각각 10%를 차지했다. β-proteobactera강은 5속 10종으로 Hydrogenophaga속이 50%, Azoarcus 20%, 나머지 Oxalicibacterium, Duganella, Xenophilus속은 각각 10%를 차지했다. γ-proteobacteria강은 13속 57종으로 Proteobacteria문에서 74%로 우점강을 차지했고, Shewanella는 23%, Pseudomonas속 12%, Cobetia 19%, Rahnella, Vibrio 및 Serratia속은 4%, 나머지 Rheinheimera, Raoultella, Pantoea, Acinetobacter, Moraxella 및 Psychrobacter속은 2%를 차지했다. Actinobacteria는 1속 2종으로 Gordonia와 Nocardioides속이 각각 50%를 나타냈다. Bacteroidetes문은 3속 5종으로 Lacihabitans, Mariniflexile속은 33%, 나머지 Dyadobacter, Cellulophaga와 Ferruginibacter속은 11%를 차지했다. Recently, Sargassum horneri, the marine weed inhabiting the shoreline, beach, and littoral sea area, has caused serious damage to intensive aquaculture farms particularly those around Jeju Island, South Korea. The purpose of this study was to investigate the diversity of microorganisms in Sargassum horneri and to provide basic data on ecological problems by identifying microbial functions. A total of 88 isolates were identified by 16S rRNA sequencing. Proteobacteria was the dominant phylum accounting for 88%, including class α-proteobacteria, six genera, and ten species. The dominating genus, Pseudobacter, accounted for 40% in Pseudorhodobacter, 20% in Paracoccus, and the remaining at 10% each were Rhizobium, Albirhodobacter, Skermanella, and Novosphingobium. Class β-proteobacteria included five genera and ten species. Genus Hydrogenophaga accounted for 50%, while genus Azoarcus accounted for 20%, and the remaining Oxalicibacterium, Duganella, and Xenophilus were 10% each. Class γ-proteobacteria with 13 genera and 57 species, accounted for 74% in phylum Proteobacteria, 23% in Shewanella, 19% in Cobetia, 12% in Pseudomonas, 4% each in Vibrio and Serratia, and 2% each in Rheinheimera, Raoultella, Pantoea, Acinetobacter, Moraxella, and Psychrobacter genera. In addition, Actinobacteria with two species of Nocardioides genera accounted for 50%, and Bacteroidetes accounted for 33%, with three genera and five species that included Lacihabitans and Mariniflexile. The remaining Dyadobacter, Cellulophaga, and Ferruginibacter genera each accounted for 11%.

프로젝트 관리의 성공에 영향을 미치는 장애요인

이아연(A-yeon Lee),문경미(Kyung-mi Moon),김승철(Seung-chul Kim) 한국산업경영시스템학회 2020 한국산업경영시스템학회지 Vol.43 No.3

As the business environment is rapidly changing with globalization and complexity of information flows, the uncertainty is also very increased for project environment. Although many studies have been conducted to find out the critical factors for project success, there still exist different views to define project success. Furthermore, implementing success formula for one project does not necessarily guarantee a success for another project since there are other elements that impede the success of project. In this regards, it is imperative to examine what are the barriers to project success. This study aims to examine the barriers that impede the success of project. Past literature was thoroughly reviewed to collect and develop a preliminary list of elements that affected project performance negatively. Experts were interviewed to refine the list and the final list of the measurement items were developed. A survey questionnaire was developed with the final list of measurement items, and a survey was conducted on the practitioners with project experience. After the survey, an exploratory factor analysis was conducted on the final list to extract the component dimensions which in turn formed the group of project barriers. The exploratory factor analysis provided ten factors, which are difficulty of process management, failure of project feasibility analysis, cost overruns and lack of cost benefits, unclarity project plan, strategic consistency error, stakeholder conflict, inaccuracy of requirement definition, disturbance of communication, technical environment change, negative attitude of top management.

Efficiency of Silver Ion-Silica Solid Phase Extraction for Elimination of Sulfur Compounds during Pesticide Multiresidue Analysis in Allium Species

Jin-Woo Park(박진우),Kyung-Mi Moon(문경미),Young-Whan Choi(최영환),Young-Guen Lee(이영근) 한국생명과학회 2010 생명과학회지 Vol.20 No.1

파속류에 다량 함유된 황화합물들은 GC/ECD에 민감하게 반응하기에, 이 농산물들의 잔류농약분석에서 방해 물질로 작용하여 농약성분의 분리정량에 어려움을 주고 있다. 최근 잔류농약분석에서 정제법으로 여러 가지 고상추출법(solid-phase extraction)이 개발되고 있으며, 본 연구에서는 황화합물을 효율적으로 제거하기 위하여, 20% 질산은(AgNO₃)용액 3 ml을 Florosil cartridge (1 g packing, 6 ml tube)에 처리한 은이온흡착 cartridge (silver nitrate impregnated florosil cartridge)를 조제하였다. 이 cartridge를 이용하여 파속류 각 2, 4, 6 및 10 g에 해당하는 추출액을 고상추출법으로 정제한 결과, 양파 6 g, 대파 4 g, 쪽파는 4 g 까지는 황화합물의 95% 이상을 제거할 수 있었다. 그리고, 40종의 농약을 각 파속류에 첨가하고 이 cartridge에서 유출하여 회수율을 분석한 결과 34종이 70~120%의 범위 안에 들었다. Since organic sulfur compounds respond to GC/ECD sensitively, they interfere with quantitative separation of pesticides during residual pesticide analysis of Allium species. In this study, it was intended to develop a rapid and simple method for pesticide multi-residues analysis through clean-up and interferences by a solid-phase extraction (SPE). An SPE method employing silver nitrate impregnated Florosil cartridge was developed and evaluated for the elimination of sulfur compounds from the test solution of Allium species during pesticide residues analysis. The silver nitrate impregnated Florosil cartridge was prepared by efflux of 3 ml of 20% silver nitrate solution through Florosil cartridge (1 g packing, 6 ml tube). The extracts equivalent to 2, 4 6, and 10 g of each sample were loaded onto the cartridge and allowed to exude, and then the exudations were analyzed by GC/ECD. More than 95% of sulfur compounds were removed from the loaded extracts equivalent, up to 6 g in onion, 4 g in spring onion and 4 g in shallot, respectively. 40 pesticides were spiked in the Allium species and loaded onto the cartridge to determine the recoveries; from this, the recoveries of 34 pesticides were within 70~120%.

빨강불가사리(Certonardoa semiregularis)에서 분리된 세균의 군집구조 분석

이해리(Hae-Ri Lee),박소현(So-Hyun Park),김동휘(Dong-Hwi Kim),문경미(Kyung-Mi Moon),허문수(Moon-Soo Heo) 한국생명과학회 2018 생명과학회지 Vol.28 No.8

불가사리를 이용하여 다양한 생리활성에 대한 연구가 많이 진행되었지만, 다른 천연물 연구에 비해 불가사리로 부터 분리한 미생물에 대한 연구는 부족하다. 이에 본 연구에서는 제주도에서 채집한 극피동물인 빨강불가사리(Certonardoa semiregularis)의 내장으로부터 Marine Agar와 R2A를 이용하여 총 103개의 균주를 분리하여 세균군집에 대해 조사하였다. 분리된 균주들은 16S rRNA유전자를 이용하여 염기서열을 분석 및 동정하였다. 그 결과 주요 계통군은 Proteobacteria (Alpha-proteobacteria 24%, Beta-proteobacteria 4%, Gamma-proteobacteria 65%) 93%, Bacteroidetes 4%, Actinobacteria 2%, Firmicutes 1%로 4개의 문이 확인되었고, 7개의 (Actinobacteria, Flavobacteria, Bacilli, Sphingobacteria, Alpha-proteobacteria, Beta-proteobacteria, Gamma-proteobacteria), 15개의 목, 19개의 과, 24속이 관찰되었다. 또한 계통 분석 결과 2개의 균주(Lysobacter sp., Pedobacter sp.)가 각각 97.55%, 97.58%로 상동성이 98% 이하로 나타나 새로운 속 또는 종으로 분류될 가능성이 있다고 여겨지며, 표준 균주와 함께 신종 후보 균주에 대한 생화학적, 형태학적 등의 추가적인 신종실험을 향후 진행해야 할 것으로 사료 된다. Although much research has focused on various bioactive substances in starfish, research on microorganisms isolated from starfish is lacking as compared with other natural products. In this study, we investigated bacterial communities in the gut of red starfish (Certonardoa semiregularis) in Jeju Island. In total, 103 bacterial strains were isolated using marine agar and R2A medium. The isolated strains were analyzed and identified using the 16S rRNA gene sequence. Based on an analysis of this gene sequence, the 103 isolated bacteria were classified into four major groups: Proteobacteria (93%: Alpha-proteobacteria, 24.8%; Beta-proteobacteria, 4%; Gammaproteobacteria, 65%) Bacteroidetes (4%), Actinobacteria (2%), and Firmicutes (1%). In addition, the isolates were divided into seven classes (Actinobacteria, Flavobacteria, Bacilli, Sphingobacteria, Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria), 15 orders, 19 families, and 24 genera. A phylogenetic analysis revealed two strains, Lysobacter sp. and Pedobacter sp., with similarity of 97.55% and 97.58%, respectively. As the similarity in the 16S rRNA gene sequence was 98% or less compared to previously identified bacteria, the two strains may possibly be classified as a new genus or species. We suggest that additional studies, including biochemical and morphological tests, should be performed to identify the new candidate strains.

Oxytetracycline의 투여방법에 따른 넙치(Paralichthys olivaceus) 체내의 잔류 특성

고창식(Chang-Sik Ko),김동휘(Dong-Hwi Kim),박소현(So-Hyun Park),문경미(Kyung-Mi Moon),허문수(Moon-Soo Heo) 한국생명과학회 2016 생명과학회지 Vol.26 No.3

어류 질병은 매년 대규모 해산물 양식 및 소비량에 따라 증가하고 있으며, 이를 예방 및 치료하기 위해 항생제인 OTC (Oxytetracycline)를 사용하고 있다. 하지만 이는 부적절한 사용 및 오남용을 하게 되면 인체 내 통증을 유발시킬 수 있다고 알려져 있다. 따라서 본 연구에서는 간이시험법 및 HPLC 분석과 함께 OTC의 약욕 처리 및 경구 투여에 따른 넙치의 피부, 간, 근육에 관해 연구하였다. 약욕처리구는 물 1톤당 25 g의 농도로 1시간 1회 실시 하였고, OTC의 투여 농도를 어체중 ㎏ 당 62.5 ㎎을 사료에 혼합시켜 7일간 경구 투여하였다. 미생물학적간이 검사법에서 약욕 처리구는 피부 직후에서만 양성을 보였고 경구 투여구에서는 77일째 양성으로 조사되었다. 그리고 근육은 14일째까지 양성을 보였다. HPLC 분석에서는 약욕 처리구인 간에서는 13일째, 피부는 37일째 잔해가 남은 것을 확인할 수 있었고 근육에서는 발견하지 못했다. 경구 투여구인 피부에서는 높은 농도(1.07 ㎎/㎏)로 나타났으며, 간에서는 소량(0.56 ㎎/㎏)을 나타냈으며 42일째의 근육에서는 남아 있지 않았다. 종합적으로 어류 양식장에서 사용 되는 OTC의 휴약 기간을 40일로 준수하면 OTC는 거의 잔류하지 않고 우리 몸에 무해하나 영양물질의 중간대사 또는 저장, 해독 등의 중요한 역할을 하는 간과 어체의 몸의 표면을 보호하고 감각작용을 하는 역할을 하는 피부는 근육에 남아 있는 OTC의 잔류량보다 몇 배의 높은 수치를 나타냈다. 하지만 이는 이전 연구에 따르면 OTC의 잔류량은 조리 온도에 따라 감소될 것으로 추측 된다. Industrial advancements have resulted in food culture development, followed by increased seafood consumption and large-scale seafood farming, which has been accompanied by an increased prevalence of fish disease. The antibiotic oxytetracycline (OTC) is commonly used to prevent and treat bacterial diseases in fish. However, overuse of OTC had led to negative aspects. In view of this, we conducted a research with regard to aspects of remnants on olive flounder skin, liver, and muscle through dipping treatment and oral feeding of OTC and analyzed the results with bioassay and HPLC quantitative analyses. The dipping treatment was carried out once with 25 g/ton/hr of OTC, and the oral treatment with 62.5 ㎎/㎏ body weight/7 days. The results underwent a bioassay analysis. The dipping group reacted only on the skin right after dipping, while the oral feeding group responded on the skin for 77 days after feeding and on the muscle for 14 days. In the dipping group, the HPLC quantitative analysis revealed remnants in the skin on the 37th day and on the 13th day in the liver group. No remnants were found in the muscle, even immediately after dipping. In the oral feeding group, there was a high concentration (1.07 ㎎/㎏) of remnant in the skin, even on the 77th day. 0.56 ㎎/㎏ in the liver, even a small amount, and no remnant in the muscle on the 42nd day. To sum up, the results suggest that it will not be harmful to our body to observe the OTC withdrawal period of 40 days with the muscle because OTC will hardly remain on it. When using olive flounder for sashimi, the skin and liver should not be used for broth, as the quantity of OTC residue is several times higher than that found in muscle. As previous studies reported that the concentration of remnants gradually decreased with heating, so it was likely to lessen, depending on the cooking temperature.

UPLC-MS/MS를 이용한 경남지역 파프리카 중 neonicotinoid계 농약 잔류 모니터링

김남국(Nam-Kuk Kim),이승화(Seung Hwa Lee),남유정(Yu Jeong Nam),문경미(Kyung Mi Moon),박민호(Min Ho Park),윤문희(Mun Hee Yun),김미영(Mi Young Kim),장현민(Hyun Min Jang),신봉식(Bong Shig Shin) 한국농약과학회 2011 농약과학회지 Vol.15 No.1