• Title/Summary/Keyword: Bacterial Diversity

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Physicochemical Properties and Microbial Analysis of Korean Solar Salt and Flower of Salt (한국산 꽃소금과 천일염의 이화학적 특성 및 미생물 분석)

  • Lee, Hye Mi;Lee, Woo Kyoung;Jin, Jung Hyun;Kim, In Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1115-1124
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    • 2013
  • The present study was conducted to ensure the diversity of domestic solar salt by analyzing the composition and microbiological characteristics of solar salt (from Docho island: DS) and the flower of salt produced in different Korean salt flats (Sinui island: SF, Bigum island: BF, and Docho island: DF). The analyses showed that the moisture content of the three types of flower of salt and solar salt ranged from 10.54~13.82% and NaCl content ranged from 78.81~84.61%. The mineral content of those salts ranged from 3.57~5.51%. The content of insoluble matter in these salts was $0.01{\pm}0.00{\sim}0.05{\pm}0.00%$. The sand content of these salts was $0.01{\pm}0.01{\sim}0.03{\pm}0.01%$. By Hunter's color value analysis, the color of the flower of salt was brighter and whiter than solar salt. The salinity of the flower of salt was a little higher than solar salt as well. The magnesium and potassium ion content of DF was $9,886.72{\pm}104.78mg/kg$ and $2,975.23{\pm}79.73mg/kg$, respectively, which was lower than the content in SF, BF, and DS. The heavy metal content of all salts was acceptable under the Korean Food Sanitation Law. The flower of salt was confirmed to be sweeter and preferable to solar salt. More than 80% of the solar salt crystals were 2~3 mm in size, whereas crystals from the flower of salt were 0.5~2 mm in size. The bacterial diversity of DF and DS were investigated by culture and denaturing gradient gel electrophoresis (DGGE) methods. The number of cultured bacteria in flower of salt was approximately three times more than solar salt. By DGGE analysis, major microbes of DF were Maritimibacter sp., Cupriavidus sp., and unculturable bacteria, and those of DS were Cupriavidus sp., Dunalidella salina and unculturable bacteria. The results of DGGE analysis showed that major microorganisms in solar salts were composed of unidentified and unculturable bacteria and only a few microorganisms were culturable.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Drug Resistance Patterns of the Bacterial Strains Isolated from Rural Areas and an Urban General Hospital (무의촌균주(無醫村菌株) 및 병원균주(病院菌株)의 항균제(抗菌劑) 내성(耐性) 양상(樣相)에 관(關)한 연구(硏究))

  • Rhee, Kwang-Ho;Kim, Ik-Sang;Shin, Hee-Sup;Cha, Chang-Yong;Lee, Seung-Hoon;Chang, Woo-Hyun;Lim, Jung-Kyoo
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.19-32
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    • 1980
  • Besides the benefits of antimicrobial agents in the control of various infectious diseases, widespread and prolonged use of particular antimicrobial agents has brought about the increase of drug-resistant strains in a community and the profound changes in the pattern of infectious diseases. In Korea, there are some remote villages where no clinics and drug stores are available and the residents in those areas are assumed to have fewer chances to contact with antimicrobial agents. In the present study, the differences in susceptibilities to 14 antimicrobial agents between the isolates from rural areas(R) and Seoul National University Hospital(SNUH, H) were studied. The isolates and their numbers were Staphylococcus aureus, R;55, H;68), Enterococci(R;28, H;30), Escherichia coli(R;40, H;40), Enterobacter aerogenes(R;25, H;21) and Klebsiella pneunoniae(R;58, H;67). Minimal inhibitory concentrations(MIC's) of penicillin, ampicillin, carbenicillin, cephalexin, tetracycline, oxytetracyline, doxycycline, minocycline, gentamicin, kanamycin, streptomycin, erythromycin, troleandomycin and co-trimoxazole were determined by agar dilution method. I. Comparison of MIC's and resistant strain proportions between isolates from SNUH and rural areas. MIC's and/or resistant strain proportions of the isolates from SNUH were significantly higher than those of the isolates from rural areas in the cases of 1. S. aureus to doxycycline, streptomycin and kanamycin. 2. E. coli to penicillin, ampicillin, carbenicillin, tetracycline, oxytetracycline, doxycycline, minocycline, streptomycin, kanamycin, erythromycin and co-trimoxazole. 3. E. aerogences to carbenicillin, tetracycline, oxytetracycline, doxycycline, minocycline, streptomycin, kanamycin, genaamicin and co-trimoxazole. 4. K pneunoniae to penicillin, ampicillin, tetracycline, oxytetracycline, doxycycline, monocycline, streptomycin, kanamycine, gentamicin and co-trimoxazole. However, the mean MIC and resistant strain proportion of S. aureus to tetracycline were higher in isolates from rural areas than in those from SNUH and Enterococci showed no differences in susceptibilities to the antimicrobial agents between isolates from rural areas and from SNUH. Therefore, in general, differenes in susceptibility to these antimicrobial agents between the isolates from rural areas and SNUH were remarkably greater and broader in gram negative enteric bacteria. II. Multiple drug resistance pattern. Patterns and incidences of multiple drug resistance were studied with penicillin, ampicillin, tetracycline, cephalexin, gentamicin, streptomcin, kanamycin and co-trimoxazole in Enterococci, E. coli, E. aeroges and K. pneumoniae. There appeared significant differences in the incidence of multiply drug-resistant strains and multiple drug resistance patterns between the isolates from SNUH and rural areas in Enterococci, E. coli, E. aerogenes and K. pneumoniae. However, there was no difference in the incidence of multiply drug-resistant strains between isolates of S. aureus from SNUH and rural areas but the pattern of multiple resistance of the SNUH strains of S. aureus was diverse, while that of the rural strains was predominantly confined to penicillin-tetracycline combination. The incidence of multigly drug-resistant strains and diversity of their patterns were the highest in E. coli strains isolated from SNUH and there were no multiply drug resistant strrains in Enterococci and K. pneumoniae strains isolated from rural areas. The number of drug-resistance determinants was also different between the isolates from rural areas and SNUH. Most of the multiply drug-resistant strains of E. coli, E. aerogenes and K. pneumoniae isolated from SNUH were resistant to more than 3 kinds of antimicrobial agents, most frequently to ampicillin, tetracycline and streptomycin, while multiply drug-resistant strains from rural areas were resistant to 2 kinds of antimicrobial agents among ampicillin, tetracycline and streptomycin. With drug-resistant E. coli strains, resistance to tetracycline which was used most widely since 1951 was most frequently involved as a part of mutliple drug-resistance, followed by resistance to ampicillin and streptomycin. This strongly suggests that emergence of drug-restant strains in a community is directly dependent on the selective pressure exerted by the antimicrobial agent used. III. Cross resistance. Cross resistance of bacteria was studied among tetracycline penicillin, aminoglycoside and macrolide derivatives by analyzing correlation coefficients of sucseptibilities using the least square method. In this study, there were high correlations among the susceptibilities to related derivatives. It appears that the relatively low correlations in susceptibilities present in some cases are due to intrinsic resistance of E. aerogenes to penicillin, Enterococci to aminoglycoside and E. coli E. aerogenes and K. pneumoniae to macrolide derivatives.

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ISOLATION AND IDENTIFICATION OF BACTERIA FROM THE ROOT CANAL OF THE TEETH DIAGNOSED AS THE ACUTE PULPITIS AND ACUTE PERIAPICAL ABSCESS (급성 치수염 및 급성 치근단 농양의 치근관으로부터의 세균 분리 및 동정)

  • Lee, Yeon-Jae;Kim, Mi-Kwang;Hwang, Ho-Keel;Kook, Joong-Ki
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.409-422
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    • 2005
  • The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 XPBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing $5\%$ sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions, This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.

Investigation of bacteria in the agricultural by-products imported for the use as media materials in mushroom cultivation (버섯재배 배지재료용 수입 농업부산물에서의 세균 조사 연구)

  • Kim, Jun Young;Kim, Susan;Kim, Seong Hwan
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.410-419
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    • 2018
  • It is urgently required to construct safety data on agricultural by-products imported for use as medium materials for domestic mushroom production. However, research on microorganisms is insufficient. This study was conducted to investigate the presence of bacteria that have the possibility of harmful effects on human, plants and mushroom in wheat straw, peatmoss, cottonseed hull, cottonseed meal, and beet pulp imported from Australia, Canada, China, Egypt, Germany. Bacteria were found in the range of $1.35{\times}10^2$ to $8.34{\times}10^6CFU/g$. As a result of 16S rDNA sequence analysis, total of 19 genera and 45 species of bacteria were identified. Bacillus genus was dominant, followed by Paenibacillus genus. At the species level, diverse species was in the order of Firmicute, Proteobacteria and Actinobacteria. Regarding the agricultural by-products, straw and peat moss had more diverse bacteria than other agricultural by-products. Among the indentified bacteria, 6 species of 5 genera (Enterobacter asburiae, Enterobacter ludwigii, Stenotrophomonas maltophilia, Pseudomonas monteilii, Bacillus anthracis, and Cellulosimicrobium funkei) were present as potent harmful bacteria to human. Surprisingly, both the human and plant pathogenic Klebsiella pneumoniae subsp. pneumonia was present. Bacillus altitudinis was present as a plant pathogen. Lysinibacillus sphaericus, an insect pathogen, and Ochrobactrum pseudogrignonense, a mushroom pathogen, were also present. The results of this study confirmed that several kinds of pathogenic bacteria were present in the agricultural by-products for the mushroom cultivation medium imported into Korea. Our work suggests that hygiene inspection and management is urgently needed for imported agricultural by-products to be safely used for mushroom production.

Distribution of Culturable Bacteria of Bioaerosol according to Land Type in Winter in the City Center (도심지 겨울철 토지피복 유형별 바이오에어로졸 중 배양성 세균 분포)

  • Kim, Jeong-Ho;Yun, Yong-Han;Kim, Hak-Gi;Lee, Myeong-Hun;Park, Yeong-jin;Lee, Dong-Jae;Sin, Yong-jin
    • Korean Journal of Environment and Ecology
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    • v.35 no.6
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    • pp.669-678
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    • 2021
  • This study surveyed three land cover types in Chungju City in Chungcheongbuk Province to check the distribution of cultured bacteria in bio-aerosols according to land cover type. It was possible to compare and analyze the distribution of bacteria according to microclimatic changes at each measurement point by examining meteorological factors and bio-aerosols according to land cover. The microclimate temperature in each measurement point was 8.7℃ for the urban forest, 10.8℃ for the waterside green area, and 10.2℃ for the urban area, indicating the urban forest had the lowest temperature among the measurement points. The relative humanity was 61.8% fin the urban forest, 59.3% in the waterside green area, and 55.7% in the urban area, indicating that the urban forest was the most humid among the measurement points. The identified bacteria were found to be 43 genera and 99 species. In terms of species diversity of cultured bacteria, 22 genera were found in the waterside green area, 21 genera in the urban forest, and 17 genera in the urban area, 37 species were found in the waterside green area, 31 species in the urban area, and 31 species in the urban forest. Bacillus toyonensis and Pseudarthrobacter oxydan were the species present in all three types of measurement sites, and Herbiconiux flava was confirmed to inhabit green areas such as urban forests and waterside green areas. The analysis result of the bacterial concentration according to the microclimatic environment in each measurement point was 333 CFU/m3 in the urban forest, 287 CFU/m3, in the waterside green area, and 173 CFU/m3 in the downtown area. The relative humidity and wind speed were analyzed to show a similar trend as the concentration. This study is expected to provide basic data for healthy urban management and green area creation by identifying the distribution of cultured bacteria in bio-aerosols according to land cover type and comparing and analyzing the traits of bio-aerosol in each measurement point.

Determination of Fire Blight Susceptibility on Wild Rosaceae Plants in Korea by Artificial Inoculation (인공접종을 통한 국내 야생 장미과 식물의 화상병 감수성 검정)

  • In Woong Park;Yu-Rim Song;Eom-Ji Oh;Yoel Kim;In Sun Hwang;Mi-Jin Jeon;Chorong Ahn;Jin-Suk Kim;Soonok Kim;Chang-Sik Oh
    • Research in Plant Disease
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    • v.29 no.1
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    • pp.23-38
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    • 2023
  • The fire blight caused by Erwinia amylovora (Ea) is a devastating disease of Rosaceae plants, including commercially important apple and pear trees. Since the first report in Korea in May 2015, it has been spreading to neighboring regions gradually. Host plants can be infected by pollinators like bees, rainfall accompanied by wind, and cultural practices such as pruning. Many studies have revealed that wild Rosaceae plants such as Cotoneaster spp., Crataegus spp., Pyracantha spp., Prunus spp., and Sorbus spp. can be reservoirs of Ea in nature. However, wild Rosaceae plants in Korea have not been examined yet whether they are susceptible to fire blight. Therefore, the susceptibility to fire blight was examined with 25 species in 10 genera of wild Rosaceae plants, which were collected during 2020-2022, by artificial inoculation. Bacterial suspension (108 cfu/ml) of Ea type strain TS3128 was inoculated artificially in flowers, leaves, stems, and fruits of each plant species, and development of disease symptoms were monitored. Moreover, the presence of Ea bacteria from inoculated samples were checked by conventional polymerase chain reaction. Total 14 species of wild Rosaceae plants showed disease symptoms of fire blight, and Ea bacteria were detected inside of inoculated plant parts. These results suggest that wild Rosaceae plants growing nearby commercial apple and pear orchards in Korea can be Ea reservoirs, and thus they should be monitored regularly to minimize the damage by Ea infection and spreading.