• Title/Summary/Keyword: Microbial profiling

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Identification and Chemotype Profiling of Fusarium Species in Korean Oat (국내 귀리의Fusarium속 균의 다양성 및 독소 화학형)

  • Choi, Jung-Hye;Nah, Ju-Young;Jin, Hyun-Suk;Lim, Su-Bin;Paek, Ji-Seon;Lee, Mi-Jeong;Jang, Ja-Yeong;Lee, Theresa;Hong, Sung Kee;Kim, Jeomsoon
    • Research in Plant Disease
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    • v.25 no.4
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    • pp.157-163
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    • 2019
  • This study aimed to assess the incidence and distribution of toxigenic fungi in Korean oat. Toxigenic fungi were isolated from oat samples collected from 12 oat fields from heading to harvest in 2017 and 2018. A total of 745 fungal colonies were isolated based on morphology and identified using marker genes. About 92% of the fungal isolates were Fusarium spp. and others were Penicillium (5.9%) and Aspergillus (2.1%). Fusarium isolates comprised mostly of F. asiaticum (83.1%), followed by F. incarnatum (5.4%), F. proliferatum (3.5%), F. fujikuroi (2.8%), F. tricinctum species complex (FTSC) 11 (1.5%) and F. graminearum (1.0%). About 97% of F. asiaticum was nivalenol type, and 3-acetyl deoxynivalenol (3.2%) and 15-acetyl deoxynivalenol (0.4%) types also were found. Pathogenicity test of the selected Fusarium isolates revealed that F. asiaticum isolates have a wide range of virulence depending on the tested plants. F. graminearum and FTSC 11 isolates from blighted spikelets were the most virulent in naked oat. All Fusarium isolates (n=18) except one (FTSC 11) produced nivalenol (0.2-7.6 ㎍/g), deoxynivalenol (0.03-6.1 ㎍/g), and zearalenone (0.1-27.0 ㎍/g) on rice medium. This study is first report that F. asiaticum causes Fusarium head blight disease of oat in Korea. These findings demonstrate the dominance of F. asiaticum in oat agroecosystems as in rice, wheat and barley in Korea.

Analysis of Microbial Composition Associated with Freshwater and Seawater

  • Lee, So-Yeon;Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.150-159
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    • 2016
  • Knowledge of the distribution and biodiversity of environmental bacteria and the ecosystem that influences them is crucial for predicting an ecosystem. However, bacterial culture methods can only analyze approximately 0.1% of the existing microorganisms, those that are readily cultured under laboratory conditions. By contrast, next-generation sequencing (NGS) has generally been known to obtain more diverse profiling of bacterial composition. We compared the bacterial communities using both a culture-dependent (MALDI-TOF) and culture-independent (NGS) methods. Environmental specimens were obtained from both freshwater and seawater. Water samples were also analyzed by both pyrosequencing and MiSeq sequencing, in order to select one NGS platform which could analyze comparatively more diverse microbiota. Bacterial distribution analyzed with MALDI-TOF showed no difference between the microbiota of freshwater and seawater, whereas the results analyzed with NGS distinguished between the two. The diversity indexes of MiSeq sequencing were higher than for Pyrosequencing. This indicated that MiSeq sequencing is capable of analyzing a comparatively wider diversity of bacteria. The genus of Flavobacterium and Planktophila were identified as being unique to freshwater, whereas EU801223 and OM43 were found in the seawater. Difference between the bacterial composition of the freshwater and seawater environments was identified by MiSeq sequencing analysis.

Reliable Identification of Bacillus cereus Group Species Using Low Mass Biomarkers by MALDI-TOF MS

  • Ha, Miyoung;Jo, Hyeon-Ju;Choi, Eun-Kyeong;Kim, Yangsun;Kim, Junsung;Cho, Hyeon-Jong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.887-896
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    • 2019
  • Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based pathogen identification relies on the ribosomal protein spectra provided in the proprietary database. Although these mass spectra can discern various pathogens at species level, the spectra-based method still has limitations in identifying closely-related microbial species. In this study, to overcome the limits of the current MALDI-TOF MS identification method using ribosomal protein spectra, we applied MALDI-TOF MS of low-mass profiling to the identification of two genetically related Bacillus species, the food-borne pathogen Bacillus cereus, and the insect pathogen Bacillus thuringiensis. The mass spectra of small molecules from 17 type strains of two bacilli were compared to the morphological, biochemical, and genetic identification methods of pathogens. The specific mass peaks in the low-mass range (m/z 500-3,000) successfully identified various closely-related strains belonging to these two reference species. The intensity profiles of the MALDI-TOF mass spectra clearly revealed the differences between the two genetically-related species at strain level. We suggest that small molecules with low molecular weight, 714.2 and 906.5 m/z can be potential mass biomarkers used for reliable identification of B. cereus and B. thuringiensis.

The Antibacterial Properties of Filtrates from Chinese Cabbage Kimchi

  • Seong-Soo CHA;JeungSun LEE;Min-Kyu KWAK
    • The Korean Journal of Food & Health Convergence
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    • v.9 no.6
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    • pp.9-19
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    • 2023
  • Lactobacillus plantarum and Leuconostoc mesenteroides are crucial functional starters and predominant isolates in a wide range of fermented foods, particularly kimchi, whose constituents exhibit bioactive properties. We previously developed a methodology using anion exchange resins to purify peptidyl compounds from Lb. plantarum LBP-K10. Antibacterial cultures of Lb. plantarum LBP-K10 were obtained from the respective cultures' supernatants and filtrates. However, conclusive evidence of the efficacy of kimchi filtrates in eradicating pathogenic bacteria is lacking. We aimed to simulate the potential effects of antibacterial filtrates that contained antibacterial compounds which were derived from cultures of Lb. plantarum LBP-K10. We acquired the kimchi filtrates using a combination of centrifugation and filtration methodologies, without the requirement for inoculation. The filtered liquid from Chinese cabbage kimchi, inoculated with Lb. plantarum LBP-K10 as a starter culture, and the non-inoculated liquid from Chinese cabbage kimchi (referred to as CCK and CCKRef, respectively) were were examined. CCK demonstrated greater inhibitory activity and a more significant bactericidal effect against the bacterial indicator strains. The minimum inhibitory concentration demonstrated comparable outcomes in tests against both Gram-positive and Gram-negative bacteria. This research offers a groundbreaking examination that displays the effectiveness of profiling peptidyl compounds within kimchi filtrates for curing bacterial infections.

Marine Metatranscriptome Profiling in the Sea Adjacent to Jeju Island, Korea, by RNA-sequencing (RNA-sequencing을 이용한 제주도 인접 바다의 메타전사체 프로파일링)

  • Hwang, Jinik;Kang, Mingyeong;Kim, Kang Eun;Jung, Seung Won;Lee, Taek-Kyun
    • Journal of Life Science
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    • v.30 no.7
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    • pp.625-629
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    • 2020
  • The Ocean is a rich source of diverse living organisms include viruses. In this study, we examined the microbial communities in the sea adjacent to Jeju Island in two seasons by metatranscriptomics. We collected and extracted total RNA, and, using the next-generation sequencing HiSeq 2000 and de novo transcriptome assembly, we identified 652,984 and 163,759 transcripts from the March and December samples, respectively. The most abundant organisms in March were bacteria, while eukaryotes were dominant in the December sample. The bacterial communities differed between the two samples, suggesting seasonal change. To identify the viruses, we searched the transcripts against a viral reference database using MegaBLAST with the most identified being bacteriophages infecting the marine bacteria. However, we also revealed an abundance of transcripts associated with diverse herpesviruses in the two transcriptomes, indicating the presence or possible threat of infection of fish in the sea around Jeju Island. This data is valuable for the study of marine microbial communities and for identifying possible viral pathogens.

Bacterial Community Profiling during the Manufacturing Process of Traditional Soybean Paste by Pyrosequencing Method (Pyrosequencing을 이용한 전통된장 제조과정 중 세균군집구조의 분석)

  • Kim, Yong-Sang;Jeong, Do-Yeon;Hwang, Young-Tae;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.275-280
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    • 2011
  • In order to evaluate the diversity and change of bacterial population during the manufacturing process of traditional soybean paste (doenjang), bacterial communities were analyzed using 16S rRNA gene-based pyrosequencing. In rice straw, the most important inoculum source for fermentation, the bacterial sequences with a relative abundance greater than 1% were assigned to four phyla, Proteobacteria (71%), Actinobacteria (20.6%), Bacteroidetes (4.2%), and Firmicutes (1.3%). Unlike bacterial community composition of rice straw, a different pattern of bacterial population in meju was observed with predominantly high abundance (99.1%) of Firmicutes. Phylum composition in young doenjang was almost same as that of meju. Major genera in young doenjang were Bacillus (81.3%), Clostridium (6.9%) and Enterococcus (6.3%) and the predominant species among bacterial population was B. amyloliquefaciens (63.6%). Abundance of the phylum Firmicutes in mature doenjang was 99.98%, which was even higher value than those in meju and young doenjang. Predominant species in mature doenjang were B. amyloliquefaciens (67.3%), B. atrophaeus (12.7%), B. methylotrophicus (6.5%), B. mojavensis (3.2%), and B. subtilis. (2.5%), which were also identified as major species of the microbial flora in meju. These results suggested that rice straw was a primary source for supplement of Bacillus species in manufacturing the traditional doenjang and that some species of Bacillus strains were mainly involved in the fermentation process of traditional doenjang.

Effects on microbial diversity of fermentation temperature (10℃ and 20℃), long-term storage at 5℃, and subsequent warming of corn silage

  • Zhou, Yiqin;Drouin, Pascal;Lafreniere, Carole
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1528-1539
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    • 2019
  • Objective: To evaluate the effects on microbial diversity and biochemical parameters of gradually increasing temperatures, from $5^{\circ}C$ to $25^{\circ}C$ on corn silage which was previously fermented at ambient or low temperature. Methods: Whole-plant corn silage was fermented in vacuum bag mini-silos at either $10^{\circ}C$ or $20^{\circ}C$ for two months and stored at $5^{\circ}C$ for two months. The mini-silos were then subjected to additional incubation from $5^{\circ}C$ to $25^{\circ}C$ in $5^{\circ}C$ increments. Bacterial and fungal diversity was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) profiling and biochemical analysis from mini-silos collected at each temperature. Results: A temperature of $10^{\circ}C$ during fermentation restricted silage fermentation compared to fermentation temperature of $20^{\circ}C$. As storage temperature increased from $5^{\circ}C$ to $25^{\circ}C$, little changes occurred in silages fermented at $20^{\circ}C$, in terms of most biochemical parameters as well as bacterial and fungal populations. However, a high number of enterobacteria and yeasts (4 to $5\;log_{10}$ colony forming unit/g fresh materials) were detected at $15^{\circ}C$ and above. PCR-DGGE profile showed that Candida humilis predominated the fungi flora. For silage fermented at $10^{\circ}C$, no significant changes were observed in most silage characteristics when temperature was increased from $5^{\circ}C$ to $20^{\circ}C$. However, above $20^{\circ}C$, silage fermentation resumed as observed from the significantly increased number of lactic acid bacteria colonies, acetic acid content, and the rapid decline in pH and water-soluble carbohydrates concentration. DGGE results showed that Lactobacillus buchneri started to dominate the bacterial flora as temperature increased from $20^{\circ}C$ to $25^{\circ}C$. Conclusion: Temperature during fermentation as well as temperature during storage modulates microorganism population development and fermentation patterns. Silage fermented at $20^{\circ}C$ indicated that these silages should have lower aerobic stability at opening because of better survival of yeasts and enterobacteria.

Microbiological Characteristics and Cytoprotective Effects of Samjung-Hwan Fermented by Lactic Acid Bacteria (유산균을 이용한 발효삼정환의 미생물 특성 및 세포 보호 효과)

  • Chang, Seju;Wang, Jing-Hua;Shin, Na Rae;Kim, Hojun
    • Journal of Korean Medicine for Obesity Research
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    • v.16 no.1
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    • pp.11-18
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    • 2016
  • Objectives: To confirm microbiological change and cytoprotective effect of Samjung-hwan (SJH) which fermented by Lactic acid bacteria from natural fermented SJH. Methods: SJH was fermented by Lactobacillus brevis and Lactococcus lactis subsp. lactis from natural fermented SJH. After 1 week of fermentation, we analysed pH and microbial profiling. We also performed measuring total polyphenol total flavonoid contents and 1,1-Diphenyl-2-picryhydrazyl (DPPH) free radical scavenging activity to investigate antioxidant ability. Cell viability was performed by using HepG2 cell. Results: pH of lactic acid bacteria inoculated group and non-inoculated group was decreased and total counts of lactic acid bateria for both group was increased after fermentation of SJH. Total polyphenol and flavonoid contents and DPPH free radical scavenging activity was increased in both group. Total polyphenol contents of lactic acid bacteria Inoculated group is more increased than non-inoculated group. HepG2 cell viability was increased in both group. Conclusions: SJH fermentd by Lactobacillus brevis and Lactococcus lactis subsp. lactis shows change in microbiological character and has cytoprotective effect. Further studies are required for investigating function of lactic acid bacteria during fermentation of SJH.

Isolation of Antimicrobial Active Substance from Usnea longissima against Sclerotial Rot (Sclerotinia sclerotiorum) (송라(Usnea longissima)추출물로부터 균핵병 병원균(Sclerotinia sclerotiorum)에 대한 항균 활성물질 탐색)

  • Kwon, Yubin;Choi, Yong-Hwa
    • Korean Journal of Organic Agriculture
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    • v.23 no.4
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    • pp.887-896
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    • 2015
  • To develop environment-friendly agricultural products with anti-microbial activity against Sclerotinia sclerotiorum as a pathogen of sclerotium disease, Usnea longissima was extracted by methanol and its extract was fractionated into several solvent fractions. The chloroform fraction, which showed the highest antimicrobial activity, was separated by silica gel-column chromatography and obtained into nine group subfractions. The nine group fractions were searched the antifungal activities by bioassay. The most active No. 3 subfraction was analyzed by GC-MS. Each mass spectra, corresponding to each peak of chromatogram, was compared to database of Wiley library. As a result, Usnic acid was identified as main compounds. In conclusion, Usnic acid isolated from Usnea longissima was antimicrobial chemical against Sclerotinia sclerotiorum as a pathogen of sclerotium disease.

Genome-wide Analysis and Control of Microbial Hosts for a High-level Production of Therapeutic Proteins

  • Kim, Sung-Geun;Park, Jung-Hwan;Lee, Tae-Hee;Kim, Myung-Dong;Seo, Jin-Ho;Lim, Hyung-Kwon
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2005.06a
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    • pp.230-232
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    • 2005
  • The formation of insoluble aggregation of the recombinant kringle fragment of human apolipoprotein(a), rhLK8, in endoplasmic reticulum was identified as the rate-limiting step in the rhLK8 secretion in Saccharomyces cerevisiae. To analyze the protein secretion pathway, some of yeast genes closely related to protein secretion was rationally selected and their oligomer DNA were arrayed on the chip. The expression profiling of these genes during the induction of rhLK8 in fermentor fed-batch cultures revealed that several foldases including pdi1 gene were up-regulated in the early induction phase, whereas protein transport-related genes were up-regulated in the late induction phase. The coexpression of pdi1 gene increased rhLK8-folding capacity. Hence, the secretion efficiency of rhLK8 in the strain overexpressing pdi1 gene increased by 2-fold comparing in its parental strain. The oligomer DNA chip arrayed with minimum number of the genes selected in this study could be generally applicable to the monitoring system for the heterologous protein secretion and expression in Saccharomyces cerevisiae. With the optimization of fed-batch culture conditions and the alteration of genetic background of host, we obtained extracellular rhLK8 at higher yields than with Pichia pastoris systems, which was a 25-fold increased secretion level of rhLK8 compared to the secretion level at the initiation of this study.

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