• Title/Summary/Keyword: High-isolation

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Natural Occurrence of Aflatoxigenic Aspergillus Species and Aflatoxins in Traditional Korean Fermentation Starters, Meju and Nuruk

  • Woo, So Young;Lee, Sang Yoo;Tian, Fei;Jeong, A-Yeong;Yoo, Cha Nee;Kang, Seung Yoon;Chun, Hyang Sook
    • Journal of Food Hygiene and Safety
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    • v.35 no.5
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    • pp.438-446
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    • 2020
  • Meju and nuruk (respectively soybean and malt) are traditional Korean fermentation starters that are vulnerable to contamination by harmful microorganisms such as aflatoxigenic fungi and their associated aflatoxins (AFs). In this study, Aspergillus spp. were isolated and identified from a total of 57 meju and 18 nuruk samples collected from Korean markets. Their potential aflatoxigenicity was investigated by examining the presence of three aflatoxin biosynthetic genes (aflO, aflP, and aflR) using multiplex polymerase chain reaction (mPCR) assays. Thereafter, aflatoxin production of isolates and the natural occurrence of AFs in meju and nuruk samples were analyzed by high-performance liquid chromatography (HPLC). A total of 177 Aspergillus isolates were identified and 130 isolates were obtained from meju samples. Of these, 25 isolates (19.2%) contained all three aflatoxin biosynthetic genes, and five (20%) of these isolates produced aflatoxins. Forty-seven of the Aspergillus isolates were obtained from nuruk samples, five of which (10.6%) expressed all three AF biosynthetic genes; however, none of these strains produced AFs. HPLC analysis showed that 88% (51/58) of the meju samples and 39% (7/18) of nuruk samples were not contaminated with AFs (below limit of detection). Among the isolates isolated from meju and nuruk, there were aflatoxigenic strains containing all three aflatoxin biosynthetic genes or producing aflatoxin in medium, but the frequency of aflatoxin contamination was low in the meju and nuruk samples.

Analysis of Potential Toxigenicity and Phylogeny using Target Genes in Aphanizomenon flos-aquae (Cyanophyceae) strains isolated from the Nakdong River (낙동강에서 분리된 Aphanizomenon flos-aquae (Cyanophyceae) 균주의 목표 유전자를 이용한 잠재적 독소 생성능 및 계통학적 분석)

  • Ryu, Hui-Seong;An, Sung-Min;Lim, Chang-Kun;Shin, Ra-Young;Park, Jong-Guen;Lee, Jung-Ho
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.137-147
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    • 2017
  • The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

The Varietal Comparison in Growth Characteristic and Yield of Vegetable Soybean (풋콩으로의 생육특성과 수량의 품종간 비교)

  • Lee, Seung-Su;Kim, Chang-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.2
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    • pp.156-160
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    • 2008
  • This study was conducted to find on the screening of a optimal variety after compare with characteristics of a colored soybean recommended and a vegetable soybean by pure line isolation from the indigenous variety in Chungnam area. The budding period in results compared with a budding property of testing varieties between the recommended colored soybean 6 varieties and the indigenous 3 varieties, was showed that it was delayed by increasing of a grain weight but the budding rate was not significant. And the harvest period was the most early day at the black colored soybean plot at the 30th July and was the latest day at the native variety (Nonsan jaerae) at the 21 th October. The stem length was showed a wide range from 25 to 104 cm, and the Asan jaerae in this result was the highest on 104 cm, but the Geomjeongolkong was the lowest on 25 cm. The number of pod per plant was searched about 9 to 111 range, and the most numerous plot in this experimental varieties was the Ilpumgeomjeongkong as 111 number, on the other hand, the least variety was the Asan jaerae as 9 number. The rate of more than 2 seed per pod in the vegetable soybean property was the highest at the Geomjeongolkong about 82%. Additionally, the Hwangkuemkong and Geomjeongkong 1 was relatively the high levels as 78% and 73%, respectively. At the pod yield per $1000\;m^2$ (10a) was the most much as 1,567 kg at the Ilpumgeomjeongkong. Also, the Sunheukkong and the Galmikong was 1,468 kg, 1,397 kg, respectively. Therefore, the Ilpumgeomjeongkong, Sunheukkong, and Galmikong were proper varieties for the vegetable cultivation.

Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocytes for Fat-Reduced Hanwoo Beef Production (한우 체지방 감소 쇠고기 생산을 위한 복강 및 피하지방 항체 개발)

  • Choi, Chang-Weon;Kim, Yu-Hyun;Kim, Sang-Jin;Song, Man-Kang;Kwon, Eung-Gi;Oh, Young-Kyoon;Hong, Seong-Koo;Choi, Seong-Ho;Baek, Kyung-Hoon
    • Food Science of Animal Resources
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    • v.28 no.5
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    • pp.651-659
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    • 2008
  • This study aimed to develop polyclonal antibodies to regional inedible adipocytes of Korean native cattle (Hanwoo) and investigate cross-reactivity of the antibodies. Patterns in plasma membrane proteins (PMPs) from abdominal and subcutaneous adipocytes of Hanwoo isolated by collagenase digestion were investigated using SDS-PAGE. As antigens, abdominal and subcutaneous adipocyte PMPs of Hanwoo were injected to sheep 3 times at 3 wk intervals for passive immunization, and non-immunized serum and antisera were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with heart, kidney, liver, lung, muscle, and spleen of Hanwoo were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes of Hanwoo were performed for analysing LDH concentration. Based on the SDS-PAGE analysis, specific proteins of PMPs in abdominal and subcutaneous adipocytes appeared despite rather similar patterns between both adipocytes. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas the antisera had relatively strong reactivity up to the level of 1:128,000 and 1:64,000 dilution. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivities of abdominal and subcutaneous adipocyte antisera were detected with PMPs of the organs. Both antisera strongly reacted with each adipocyte PMPs and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and have safety in cross-reactivities with body organs. Further studies on in vivo cross-reactivity and fat reduction of the antibodies against abdominal and subcutaneous adipocytes PMPs of Hanwoo should be required for inedible fat-reduced high quality beef production.

Effect of Noise in Human Body (소음이 인체에 미치는 영향)

  • 이영노
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1972.03a
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    • pp.7-8
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    • 1972
  • The effects of noise exposure are of two types: Nonauditory effects and auditory effects. Nonauditory effects of noise exposure are interference with communication by speech, sleeping and emotional behavior. The noise will cause the high blood pressure and rapid pulse, also that decrease the salivation and gastric juice. in experimentaly showed that the Corticoid hormon: Gonatotropic hormone were decrease and Thyrotropic hormoone is increase. Auditory effect of noise exposure. when the normal ear is exposed to noise at noise at hamful intensities (above 90㏈) for sufficiently long periods of time, a temoral depression of hearing results, disappearing after minutes or hours of rest. When the exposure longer or intesity greater is reached the Permanent threshold shift called noise-induced hearing loss. Hearing loss resulting from noise exposure presents legal as well as medical problems. The otologist who examines and evaluates the industrial hearing loss cases must be properly informed, not only concerning the otologic but also about the physical and legal aspects of the problems. The measurement of hearing ability is the most important part of a hearing conservation, both preplacement and periodic follow-up tests of hearing. The ideal hearing conservation program would be able to reduce or eliminate the hazardous noise at its source or by acoustic isolation of noisy working area and two ear protections (plugs and muff type) were developed for personal protection.

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Microflora and enzyme activity of conventional Meju, and isolation of useful mould (재래식 메주의 미생물군, 효소역가 및 유용 균주의 분리)

  • Choi, Seong Hyun;Lee, Mi Hyun;Lee, Seuk Keun;Oh, Man Jin
    • Korean Journal of Agricultural Science
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    • v.22 no.2
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    • pp.188-196
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    • 1995
  • To obtain useful mould strain in soybean fermentation industry, we collected conventional Meju all over the Korea and tested existance of aflatoxin in Meju collected. Also, we measured distribution of microorganism and enzyme activity of Meju and isolated Aspergillus oryzae O4-5 as a industrially useful mould. The results obtained were summarized as follows: 1. Aflatoxin was not detected by EZ-Screen Test Kit and HPLC analysis in various Meju sample collected all over the Korea from 1994.12 to 1995.2. 2. Colony forming units(CFU) of mould, yeasts, aerobe, and anaerobe were $1.3{\times}10^4-2.8{\times}10^6$, $1{\times}10^2-1.5{\times}0^6$, $2.0{\times}10^7-8.0{\times}10^8$ and $3.0{\times}10^6-7.3{\times}10^8$ per gram of Meju, respectively, indicating that CFU inter Meju samples were varied with big difference. 3. The activities of ${\alpha}$-amylase and gluco-amylase were 5-80 units and 2-34 units per g of Meju, respectively. It was shown that enzyme activity was varied depending on where the Meju was collected. 4. The activities of acidic, neutral and alkaline protease were 5-33 units, 5-302 units and 5-363 units per g of Meju, respectively. Acidic protease of Improved Meju made by D-Company was higher than that of conventional Meju as 66 units per g of Meju. 5. CNU O4-5 strain selected as a noble strain could produce amylase and protease in high level, and identified as a strain that belongs to Aspergillus oryzae. 6. The activities of acidic and neutral protease of Aspergillus oryzae CNU O4-5 strain isolated were about 10% higher than those of Aspergillus oryzae JM which has been used in soybean fermentation industry. Amylase activity of CNU O4-5 strain was similar to Aspergillus oryzae JM.

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Isolation, Quality Evaluation, and Seasonal Changes of Bakkenolide B in Petasites japonicus by HPLC (머위로부터 Bakkenolide B의 순수분리, HPLC분석 방법 및 채취 시기별 함량 분석)

  • Kim, Tae Hoon;Kim, Do Youn;Jung, Won Jung;Nagaiya, Ravichandran;Son, Beung Gu;Park, Young Hoon;Kang, Jum Soon;Lee, Young Jae;Im, Dong-Soon;Lee, Young-Geun;Choi, Yung Hyun;Choi, Young-Whan
    • Journal of Life Science
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    • v.24 no.3
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    • pp.252-259
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    • 2014
  • The leaves of Peatasites japonicus are a traditional oriental medicine with diverse biological activities. A simple and specific analytical method for the quantitative determination of bakkenolide B constituents from methanolic extract of the leaves of P. japonicus was developed. Bakkenolide B was isolated from the leaves of P. japonicus, and its structure was elucidated based on 1D, 2D NMR, and GC-MS spectral data. A liquid chromatographic method was developed to evaluate the quality of P. japonicus through determination of major active compound, bakkenolide B. The wavelengths at 254 and 215 nm were chosen to determine bakkenolide B. The recovery of the method was in the range of 98.6 to 103.1%, and bakkenolide B showed good linearity ($r^2$=0.999) within test ranges. The developed method was applied to the determination of bakkenolide B in the plant part and seasonal changes. The results showed that the content of bakkenolide B in the leaf was higher than in the petiole and rhizome. In this study, a simple, rapid, and reliable high-performance liquid chromatography method was used to determine the percentage and composition of bakkenolide B in P. japonicus procured from different Petasites species plants in South Korea. The method can be employed in routine quantitative analysis and quality control of different products in the market.

Application of a New Conjugation Method to Fish Pathogenic Bacteria Containing R Plasmid for the Analysis of Drug-Resistant Status in Aquaculture (새로운 conjugation 방법을 응용한 R plasmid 함유 어병세균의 분리와 양식장 내성균의 현황 분석)

  • Yoo Min Ho;Jeong Joon Beom;Kim Eun Heui;Lee Hyoung Ho;Jeong Hun Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.2
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    • pp.115-121
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    • 2002
  • To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

Potential probiotics activity of Bacillus spp. from traditional soybean pastes and fermentation characteristics of Cheonggukjang (전통장류유래 Bacillus spp.의 프로바이오틱스 활성과 청국장 발효 특성)

  • Ryu, Myeong Seon;Yang, Hee-Jong;Kim, Jin Won;Jeong, Su-Ji;Jeong, Seong-Yeop;Eom, Jeong-Seon;Jeong, Do-Youn
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1168-1179
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    • 2017
  • In this study, we tried to screen the Bacillus strain having safety probability by isolation of strains from traditional fermented food, measurement of probiotic properties, and the fermentative characteristics of Cheonggukjang. We isolated 400 Bacillus-like isolates from traditional fermented foods. Selected strains examined on the prevalent characteristic such as extracellular enzyme and antibacterial activities, and their safety probability was confirmed by biogenic amine productivity, hemolytic, and harmful substances and enzyme productivity. We selected the 5 strains by analysis of biogenic amine, antibacterial and B. cereus toxic associated gene. Five selected strains were examined on cell surface hydrophobicity, and bile and acid tolerance, and we selected the SRCM100730 as the final strain. SRCM100730 was confirmed B. amyloliquefaciens by 16S rRNA sequencing, and named the B. amyloloquefaciens SRCM100730 (KCCM11966P). Finally, we manufactured Cheonggukjang using SRCM100730 for confirmation of fermentation properties. Manufactured Cheonggukjang did not contain B. cereus, and showed that ${\gamma}$-PGA and extracellular enzyme activities were superior to commercial Chunggukjang. Amino nitrogen content was 544.02 mg% and 26 free amino acid were detected, and the bitterness-related amino acid content was lower than commercial Cheonggukjang. Especially, the amount of GABA was 3 fold higher than commercial Cheonggukjang. These results suggest that SRCM100730 have high availability in commercial probiotics market and fermented food industry.

Isolation and Characteristics of a Phenol-degrading Bacterium, Rhodococcus pyridinovorans P21 (페놀분해세균 Rhodococcus pyridinovorans P21의 분리 및 페놀분해 특성)

  • Cho, Kwang-Sik;Lee, Sang-Mee;Shin, Myung-Jae;Park, Soo-Yun;Lee, Ye-Ram;Jang, Eun-Young;Son, Hong-Joo
    • Journal of Life Science
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    • v.24 no.9
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    • pp.988-994
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    • 2014
  • The effluents of chemical and petroleum industries often contain non-biodegradable aromatic compounds, with phenol being one of the major organic pollutants present among a wide variety of highly toxic organic chemicals. Phenol is toxic upon ingestion, contact, or inhalation, and it is lethal to fish even at concentrations as low as 0.005 ppm. Phenol biodegradation has been studied in detail using bacterial strains. However, these microorganisms suffer from substrate inhibition at high concentrations of phenol, whereby growth is inhibited. A phenol-degrading bacterium, P21, was isolated from oil-contaminated soil. The phenotypic characteristics and a phylogenetic analysis indicated the close relationship of strain P21 to Rhodococcus pyridinovorans. Phenol biodegradation by strain P21 was studied under shaking condition. The optimal conditions for phenol biodegradation by strain P21 were 0.09% $KNO_3$, 0.1% $K_2HPO_4$, 0.3% $NaH_2PO_4$, 0.015% $MgSO_4{\cdot}7H_2O$, 0.001% $FeSO_4{\cdot}7H_2O$, initial pH 9, and $20-30^{\circ}C$, respectively. When 1,000 ppm of phenol was added to the optimal medium, the strain P21 completely degraded it within two days. Rhodococcus pyridinovorans P21 could grow in up to 1,500 ppm of phenol as the sole carbon source in a batch culture, but it could not grow in a medium containing above 2,000 ppm. Moreover, strain P21 could utilize toxic compounds, such as toluene, xylene, and hexane, as a sole carbon source. However, no growth was detected on chloroform.