• 제목/요약/키워드: Isolation System

검색결과 1,762건 처리시간 0.028초

Characterisation of fungal contamination sources for use in quality management of cheese production farms in Korea

  • Kandasamy, Sujatha;Park, Won Seo;Yoo, Jayeon;Yun, Jeonghee;Kang, Han Byul;Seol, Kuk-Hwan;Oh, Mi-Hwa;Ham, Jun Sang
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제33권6호
    • /
    • pp.1002-1011
    • /
    • 2020
  • Objective: This study was conducted to determine the composition and diversity of the fungal flora at various control points in cheese ripening rooms of 10 dairy farms from six different provinces in the Republic of Korea. Methods: Floor, wall, cheese board, room air, cheese rind and core were sampled from cheese ripening rooms of ten different dairy farms. The molds were enumerated using YM petrifilm, while isolation was done on yeast extract glucose chloramphenicol agar plates. Morphologically distinct isolates were identified using sequencing of internal transcribed spacer region. Results: The fungal counts in 8 out of 10 dairy farms were out of acceptable range, as per hazard analysis critical control point regulation. A total of 986 fungal isolates identified and assigned to the phyla Ascomycota (14 genera) and Basidiomycota (3 genera). Of these Penicillium, Aspergillus, and Cladosporium were the most diverse and predominant. The cheese ripening rooms was overrepresented in 9 farms by Penicillium (76%), while Aspergillus in a single farm. Among 39 species, the prominent members were Penicillium commune, P. oxalicum, P. echinulatum, and Aspergillus versicolor. Most of the mold species detected on surfaces were the same found in the indoor air of cheese ripening rooms. Conclusion: The environment of cheese ripening rooms persuades a favourable niche for mold growth. The fungal diversity in the dairy farms were greatly influenced by several factors (exterior atmosphere, working personnel etc.,) and their proportion varied from one to another. Proper management of hygienic and production practices and air filtration system would be effective to eradicate contamination in cheese processing industries.

Laser Captured Microdissection을 이용한 유전자 발현에 대한 연구 (I): RT-PCR을 위한 난자의 RNA 추출 및 증폭을 위한 최소한도의 확립 (Analysis of the Gene Expression by Laser Captured Microdissection (I): Minimum Conditions Required for the RNA Extraction from Oocytes and Amplification for RT-PCR)

  • 박창은;고정재;차광렬;이경아
    • Clinical and Experimental Reproductive Medicine
    • /
    • 제28권3호
    • /
    • pp.183-190
    • /
    • 2001
  • Objective: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. Methods : Mouse ovaries were fixed and cut into serial sections (7 im thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell $II^{TM}$ system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). Results: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GA PDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 $\grave{i}$ l with 50 oocytes, thus the resting 19.75 il cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. Conclusion: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.

  • PDF

야생꿩으로부터 가금티프스 억제균 Sphingomonas sanguis의 선발 및 항Salmonella 물질 생산 조건 (Isolation and Identification of Sphingomonas sanguis from Wild Pheasant and Production of Antagonistic Substance against Fowl Typhoid causing Salmonella gallinarum)

  • 류향선;이현승;임종희;김진락;김상달
    • Applied Biological Chemistry
    • /
    • 제47권1호
    • /
    • pp.27-32
    • /
    • 2004
  • 가금티프스는 닭, 오리, 칠면조, 꿩 등 가금류에 Salmonella gallinarum이 원인균이 되어 발병하는 질병이다. 본 연구는 가금티프스를 방제하기 위한 생균제 개발을 위한 기초조사의 목적으로 가금티프스 원인균인 5. gallinarum 의 생육을 저해시킬 수 있는 길항균을 야생꿩에서 분리, 선발했다. 최종적으로 3종의 우수 길항균주를 선발하였으며 선발, 분리된 길항균을 동정한 결과 Ll9, L33균주는 similarity가 100%인 Sphinromonas sanguis로 동정 되었고 L50균주는 similarity가 96.2%로써 Sphinromonas sanguis의 근연종으로 동정되었다. 선발된 길항균주가 생산한 길항물질의 특성을 확인해본 결과, S. gallinarum을 저해하는 물질이 고분자성 물질이고 내열성임을 추정 할 수가 있었다. S. sanguis Ll9의 경우 배양 20시간 이후부터 길항물질이 생산되었으며 탄소원, 질소원을 각각 maltose, $(NH_4)_2SO_4$를 첨가하였을 때 가장 높은 길항력을 나타내었다. S. sanguis L33의 경우 배양 22시간 이후 부터 길항물질이 생산되었으며 탄소원, 질소원을 각각 galactose, urea를 첨가하였을 때 가장 높은 길항력을 나타내었다. S. sanguis L50은 배양 10시간 이후부터 길항물질을 생산하였으며 탄소원, 질소원을 각각 saccharose, $(NH_4)_2S_2O_8$음로 하였을 때 가장 높은 길항력을 나타내었다.

메기의 껍질로부터 항균성 펩타이드의 정제 (Purification of Antibacterial Peptide from the Skin of the Catfish Silurus asotus)

  • 손희영;고혜진;박남규
    • 생명과학회지
    • /
    • 제26권3호
    • /
    • pp.296-301
    • /
    • 2016
  • 메기(Silurus asotus)의 껍질로부터 항균성 펩타이드를 정제하였다. 메기 껍질의 산 추출물은 단계적 농도구배조건으로 Sep-Pak C18에 의해 부분적으로 정제되었으며, 그 중에서 특히 60% 메탄올 분획(RM60)이 Escherichia coli D31에 대해 가장 좋은 항균활성을 나타내었다. 이 RM60을 사용하여 이온교환 및 5단계의 연속적인 역상 HPLC로 정제하였다. 정제된 펩타이드의 분자량과 아미노산 서열분석은 MALDI-TOF MS와 에드만 분해법으로 분석하였다. 이 펩타이드의 분자량은 약 4182.1 [M+H]+이었으며, 분석된 이 물질의 부분적인 일차구조서열은 다음과 같다; PALXXKARREAKVKF. 이러한 결과는 메기의 껍질에서 존재하는 이 펩타이드가 메기의 껍질에서 반응하는 선천성 방어 시스템에서 중요한 역할을 하고 있다고 여겨진다.

전염병관리 관련법령의 변화 추이분석 및 향후 개정방향에 관한 연구 (The amendment tendency analysis of the Korean Infectious Disease Prevention Act and a recommendation for the next amendment)

  • 황창용;오희철;이덕형;박기동;이종구
    • Journal of Preventive Medicine and Public Health
    • /
    • 제31권3호
    • /
    • pp.540-563
    • /
    • 1998
  • This Study has been carried out to make a recommendation for the next amendment of the Infectious Disease Prevention Act with a specific focus on the kind of notifyable disease. Korean, Japanese, German, U.S, English and French acts on infectious diseases prevention were reviewed, compared with and analized in regards of numbers and kinds of notifyable infectious diseases and their tendency of amendments. An criteria was designed to assess the level of validity of diseases to be designated in the act. Four items, the fatality (greater than 10% or not), the possibility to make a big epidemic, the availability of efficient vaccination and the usefulness of isolation, are used in the assessment. This index is applied to the diseases in Korean and other countries' Infectious Disease Prevention Acts. Results are as follows: 1. The Korean Infectious Disease Preventon Act has a unique way of classifying the notifyable infectious disease, that is, the first, the second and the third class. But the author cannot find the basis of classification. No other countries reviewed have the similar classification. 2. The ten diseases, cholera, plague, yellow fever, diphtheria, typhoid fever, poliomyelitis, rabies, tetanus, malaria, and meningococcal meningitis are designated as the notifyable diseases not only in Korea but also in Japan, Germany, United States, England and france. 3. Thirty seven diseases including small pox, Lassa fever, anthrax, influenza, German measles, Legionellosis, infection with E. coli O157:H7, Q-fever, brucellosis, Lyme disease are designated as legal disease at least one of the above mentioned countries. 4. The Korea has been coped with the change of the infectious disease occurrence for last fifty years in amendment of the Infectious Disease Prevention Act. 5. Japan has a special infectious surveillance system composed of 3,880 clinics throughout the whole country. 6. Germany has classified infectious diseases in five categories which are based on seriousness of disease. Any confirmed death, cases and suspected cases in class I should be reported within 24 hours. But only confirmed death and cases in class II, but not suspected cases, are reportable in Germarny. 7. Plague, bacillary dysentery, pertussis, mumps, Japanese encephaltis and Korean hemorrhagic fevers are diseases with high credits validity index among Korean legal disease. 8. German measles, anthrax, E. coli O157 : H7 infection, Lassa fever, Q-fever, brucellosis are high in validity index among those which are not designated in Korea but designated in other countries. In conclusion, the Korean Infectious Disease Prevention Act has well been coped with the changes of infectious disease occurrence for last fifty years, but the classification basis and the validity of diseases to be designated as legal diseases is worth reevaluating.

  • PDF

Activin-A 처리에 의해 분화 촉진된 인간 배아 줄기세포 유래 내배엽성 세포의 효과적인 정제 (Effective Isolation of Endodermal Lineage Cells Derived from Human Embryonic Stem Cells Post Activin-A Treatment)

  • 김문규;문성환;박순정;이경일;신정민;장재우;정형민
    • Reproductive and Developmental Biology
    • /
    • 제34권3호
    • /
    • pp.135-141
    • /
    • 2010
  • Embryoid bodies (EBs) generated from human embryonic stem cells (hESCs) include spontaneously induced endodermal lineage cells (ELCs). Activin-A plays important roles in the endoderm differentiation of hESCs. Despite studies on the generation of ELCs from hESCs with treatment of Actvin-A, it was unclear for localization and pattern of ELCs by Activin-A during differentiation of hESCs. Accordingly in this study, we knew that Actvin-A increased the cystic EBs formation, including the highly enriched AFP (endoderm lineage specific marker)-expressing cells in the surface of cystic EBs. To induce the EBs formation from undifferentiated hESCs, cells were transferred onto petri-dish and cultured in suspension condition without bFGF removed hESC media (EB media) for 3 days. Next to investigate the effect of Activin-A, EBs were subsequently cultured in EB media supplement with 100 ng/ml Activin-A for 3 days. After 5~7 days of Activin-A treatment, cystic EBs began to appear which increased in numbers reaching ~60% of initially formed EBs over 5 days. Endoderm lineage marker, AFP were highly expressed and specifically localized at the surface region of cystic EBs comparison with normal EBs. We next attached the cystic EBs onto gelatin-coated plates and cultured for 5 days. In the results of real-time PCR and immunocytochemistry analysis, AFP-expressing cells migrated and localized at the outgrowth region of attached cystic EBs. To obtain the AFP-expressing cells of the outgrowth region, we manually isolated by using micro-dissection and cultured them. These cells strongly express AFP over 70% of isolated cells post re-plating. Here, we first showed an expression pattern of specifically localized ELCs by Activin-A during differentiation of hESCs. From this observation, we could highly purified ELCs from undifferentiated hESCs. Taken together, our system will provide a novel and efficient option to generate ELCs from hESCs.

ICNP의 후향적 개발방법에 의한 한국가족현상 (Development of Family Nursing Phenomena in Korea by Retrospective Method of ICNP)

  • 윤순녕;김현숙;권영숙;박경민;김화중;이지현;고영애;소애영;양순옥;전경자;이인숙;김염임;김은희
    • 지역사회간호학회지
    • /
    • 제10권2호
    • /
    • pp.275-290
    • /
    • 1999
  • The Objectives of this study were to identify family nursing phenomena at the community in Korea and to contribute to build up family domain of International Classification for Nursing Practice. The method of this study was used retrospective one among three methods to develop ICNP during the period from April 1997 to June 1999. The procedure was to choose nursing phenomena using preliminary terms(stepl) from the reports on family nursing care of the nursing students of 5 junior colleges of nursing and 5 colleges of nursing. The study group members identified 3 common family nursing phenomena with 5 characteristics related to each phenomenon. In order to consensus the appropriate characteristics of a phenomenon(step2), 17 study group members had regrouped nursing phenomena and scored its characteristics 5 times. The essential characteristics of each family phenomenon were selected above 3.5 mean score from related characteristics(step 3). Finally, 17 phenomena were named preferred terms such as following, that was selected after investigated preliminary terms(step4). Family nursing phenomena in Korea are named as Lack of family interaction in community. Social isolation. Lack of social support system in community. Disturbance in parent role, Disturbance in marital role, Dissatisfaction of sexual life, Disturbance in family communication, Inappropriate family coping, Lack of family intimacy, Inappropriate family power structure, Family violence. Unhealthy life style. Deficit of financial management skill and support. Inadequate care a sick member. Insecure safety and hygiene in neighborhood, Inadequate home-sanitation. Inadequate home-making. Family nursing phenomena in Korea were partially confirmed family architecture of ICNP, Beta version. by this study. Further study on Family nursing phenomena in Korea will be required to support evidence through literature review of nursing classifications or field studies.

  • PDF

암모니아 입하 및 저장시설에서의 위험도 관리 (Risk Management for Ammonia Unloading and Storage Tank Facility)

  • 정윤서;우인성;임종우
    • 한국가스학회지
    • /
    • 제21권5호
    • /
    • pp.95-103
    • /
    • 2017
  • 제조산업 현장에서는 수많은 유해위험물질이 사용, 처리 및 생산되고 있으며, 암모니아를 취급하는 시설에서 많은 누출, 화재 폭발사고가 보고되고 있다. 유해위험시설에서의 위험관리 및 사고예방을 위하여 공정안전관리(PSM), 가스안전관리(SMS), 장외영향평가(ORA) 제도 등 다양한 안전환경관리 프로그램이 국내 산업현장에 적용되고 있고 학계에서도 위험성평가 관련 연구가 활발하게 진행되고 있다. 본 연구에서는 암모니아 입하 및 저장 시설을 대상으로 정량적 위험성평가가 수행되었는데 사고 시나리오에 대한 피해영향범위 산정에는 장외영향평가용 KORA 프로그램, 사고 빈도 분석에는 LOPA PFD 활용 방식을 적용하였다. 평가 결과 추정된 위험도를 완화하고 지속적인 위험을 관리하는 방안으로 누출 감지 및 비상 차단, 물 분무 및 증기 희석 설비, 방류벽 및 트렌치, 누출 비산 방호 등 공정 안전설계 하드웨어 개선부분과 위험관리기준 효과적 적용, LOPA 보완적용, 보조 피해영향범위 산정 프로그램 활용, 위험도기반 공정안전관리제도 적용, 공정위험성 재평가 및 이행성 관리 등 프로그램 및 제도 운용 측면에서의 방안이 제시되었다.

Isolation and Development of Quantification Method for Cyanidin-3-Glucoside and Cyanidin-3-Rutinoside in Mulberry Fruit by High-Performance Countercurrent Chromatography and High-Performance Liquid Chromatography

  • Choi, Soo-Jung;Jeon, Heejin;Lee, Chang Uk;Yoon, Shin Hee;Bae, Soo Kyung;Chin, Young-Won;Yoon, Kee Dong
    • Natural Product Sciences
    • /
    • 제21권1호
    • /
    • pp.20-24
    • /
    • 2015
  • Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column ($4.6{\times}250mm$ I.D., $5{\mu}m$) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with $r^2=0.9996$) in the concentration range $15.625-500{\mu}g/mL$, and the relative standard deviations (RSD%) of intra- and inter-day variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.

혐기성 상태에서 암모니움 이온과 질산성 질소를 제거하는 미생물의 분리 및 배양조건 (Isolation and Culture Conditions of a Pseudomonas Strain Capable of Removing $NH_4^+\;and\; NO_3^-$ Simultaneously in Anaerobic Conditions)

  • 김영주;송영채;김종오;박흥석
    • 한국미생물·생명공학회지
    • /
    • 제33권1호
    • /
    • pp.65-69
    • /
    • 2005
  • $NH_4NO_3$을 유일한 질소원으로 하는 배지를 제작하여 혐기적 조건에서 P. aeruginosa AE-1-3을 분리하였다. 이 균은 $1.0\%$ glucose를 배지에 첨가한 조건에서 $0.1\%\;NH_4NO_3$를 완전히 제거하였다. P. aeruginosa AE-1-3은 $0.5\%$ glucose존재 하에서도 $0.1\%\;NH_4NO_3$를 완전히 제거하였으며 균의 활성을 높이기 위해 배지에 $Cu^{2+},\;Sn^{2+},\;Zn^{2+}$를 첨가함으로써 $0.3\%\;NH_4NO_3$도 제거할 수 있었다. 본 균의 $NH_4NO_3$거시의 유기물 이용경로를 알아보기 위해 COD를 측정한 결과 $0.1\%\;NH_4NO_3$$NO_3^-$가 완전히 제거되고 $NH_4^+$$50\%$제거되는 동안 glucose를 전혀 필요로 하지 않는 결과를 확인했다. 이 결과로 본 균이 heterotrophy 이지만 혐기성 및 무유기물 조건에서 $NO_3^-$$NH_4^+$를 제거한다는 것을 알 수 있다. P. aeruginosa AE-1-3은 autotrophy보다 높은 생육과 높은 $NH_4^+$$NO_3^-$제거 능력을 보여주었다. 본 균은 저 유기물상태에서 고농도 $NH_4^+$$NO_3^-$를 완전히 제거할 수 있는 가능성을 보여 주고 있다. 본 균은 우리나라처럼 C/N비가 낮은 축산폐수 및 하$\cdot$폐수를 처리하는데 응용될 수 있다면 폐수 속의 유기물만으로도 수중에서 질소를 효과적으로 제거할 수 있는 방법이 될 수 있을 것으로 기대된다.