• Title/Summary/Keyword: Bacterial 16S rRNA sequencing

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Lactic Acid Fermentation of Gamju Manufactured Using Medicinal Herb Decoction (약초 추출액을 사용하여 제조한 감주의 젖산발효)

  • Cho, Kye-Man;Ahn, Byung-Yong;Seo, Weon-Taek
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.649-655
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    • 2008
  • In this study, the characteristics of the lactic fermentation of gamju manufactured using a medicinal herb decoction were assessed. A bacterial strain, LAB19, which is used for the induction of lactic fermentation into gamju, was isolated from dried persimmon and identified as Leuconostoc mesenteroides on the basis of morphological, physiological, and chemotaxonomical features, and 16S rRNA sequencing analysis. After 60 hours of lactic fermentation with Leuconostoc mesenteroides LAB19 at $25^{\circ}C$, the gamju was determined to contain 141.3 g/L of reducing sugar, 5.33 g/L of acids, and 1.19 g/L of soluble phenolics. Approximately 90% of reducing sugar and 58% of acids were maltose and lactic acid, respectively. Free radical scavenging activities were retained at levels between 76.6 to 75.7% during the lactic fermentation of gamju.

The Infection Characteristics of Vibrio scophthalmi Isolated from Olive Flounder, Paralichthys olivaceus (양식 넙치, Paralichthys olivaceus에서 분리한 Vibrio scophthalmi의 감염 특성)

  • Kim, Su Hyun;Woo, Sung Ho;Lee, So Jung;Park, Soo Il
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.207-217
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    • 2013
  • Recently high mortality of cultured olive flounder, Paralichthys olivaceus occurred frequently at the fish farms in Ulsan, Korea. The diseased fish showed skinny body and swimming behavior around the water surface with liver atrophy and white enteritis as internal signs. The isolated bacteria were identified to V. scophthalmi by biochemical test, nucleotide analysis of 16S rRNA and dnaJ gene sequencing. The pathogen of this study showed strong pathogenicity as 75% mortality to olive flounder by intraperitoneal injection of $1{\times}10^6$ CFU/fish. The pathological sign was not different between the naturally diseased fish and the artificially infected fish. Histopathological changes were shown to liver atrophy, desquamation of the intestinal mucosa and hyaline droplet like as other previous studies.

Isolation of Tetracycline-resistant Lactic Acid Bacteria from Kimchi (김치에서 tetracycline 내성 유산균의 분리)

  • Kang, Hyo-Jin;Kim, Byung-Chun;Park, Wan
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.1-6
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    • 2004
  • Tetracycline resistant bacterial strains were isolated from 10 batches of Kimchi among 50 batches collected in Taegu restrict. The MIC of tetracycline ranged between 25 and> 100 ㎖/l. Total genomic DNA preparation from all 10 tetracycline resistant lactic acid bacterial isolates were subjected to PCR amplification with class-specific primers for tet(M) and tet(O). In only one isolate, HJ9, tet(M) was detected. By Southern blotting and hybridization with a tet(M)-specific probe, the tet(M) gene of HJ9 isolate could be localized on a plasmid. The partial nucleotide sequence and deduced amino acid sequence of tet(M) of HJ9 showed 90-99% and 94-100% homology to those of Gram positive bacteria, respectively. With sequencing of 16S rRNA, HJ9 isolate from Kimchi was identified as Lactobacillus sakei. From these results, Kimchi can be considered potential vehicle for the spread of antibiotic-resistant lactic acid bacteria along the food chain to the consumer.

Comparison Analysis of Swine Gut Microbiota between Landrace and Yorkshire at Various Growth Stages (두 돼지 종의 다양한 성장단계에 따른 장내미생물 비교분석)

  • Unno, Tatsuya
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.308-312
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    • 2014
  • In this study, we conducted a next generation sequencing based microbial community analysis to investigate gut microbiota of the two commercially most available swine breeds, Yorkshire and Landrace. Bacterial 16S rRNA gene was amplified from fecal DNA using universal primer sets designed for V4 regions. Our comparison analysis of the gut microbiota of the two breeds suggested that their gut microbiota changed depending on the growth stages, while the difference between the two breeds was insignificant. However, there was a limited number of genera, the abundance of which was found to be different between the breeds. Those included the genus Xylanibacter in the Yorkshire samples, which was previously reported as a fiber digesting bacteria, likely increasing energy harvesting capacity of swine. In addition, others included opportunistic pathogens mostly found in the Yorkshire samples while the Landrace samples had significantly more prevalent Clostridium_IV species that were known to play a key role in systemic immunity of hosts. While microbial community shifts was found to be associated with growth stages, the difference between the two breeds seemed to be insignificant. However, there were several bacterial genera showing differential abundance, which may affect growth of hosts.

Exploring the Potentiality of Novel Rhizospheric Bacterial Strains against the Rice Blast Fungus Magnaporthe oryzae

  • Amruta, Narayanappa;Kumar, M.K. Prasanna;Puneeth, M.E.;Sarika, Gowdiperu;Kandikattu, Hemanth Kumar;Vishwanath, K.;Narayanaswamy, Sonnappa
    • The Plant Pathology Journal
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    • v.34 no.2
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    • pp.126-138
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    • 2018
  • Rice blast caused by Magnaporthe oryzae is a major disease. In the present study, we aimed to identify and evaluate the novel bacterial isolates from rice rhizosphere for biocontrol of M. oryzae pathogen. Sixty bacterial strains from the rice plant's rhizosphere were tested for their biocontrol activity against M. oryzae under in vitro and in vivo. Among them, B. amyloliquefaciens had significant high activity against the pathogen. The least disease severity and highest germination were recorded in seeds treated with B. amyloliquefaciens UASBR9 (0.96 and 98.00%) compared to untreated control (3.43 and 95.00%, respectively) under in vivo condition. These isolates had high activity of enzymes in relation to growth promoting activity upon challenge inoculation of the pathogen. The potential strains were identified based on 16S rRNA gene sequencing and dominance of these particular genes were associated in Bacillus strains. These strains were also confirmed for the presence of antimicrobial peptide biosynthetic genes viz., srfAA (surfactin), fenD (fengycin), spaS (subtilin), and ituC (iturin) related to secondary metabolite production (e.g., AMPs). Overall, the results suggested that application of potential bacterial strains like B. amyloliquefaciens UASBR9 not only helps in control of the biological suppression of one of the most devastating rice pathogens, M. grisea but also increases plant growth along with a reduction in application of toxic chemical pesticides.

Universal Indicators for Oil and Gas Prospecting Based on Bacterial Communities Shaped by Light-Hydrocarbon Microseepage in China

  • Deng, Chunping;Yu, Xuejian;Yang, Jinshui;Li, Baozhen;Sun, Weilin;Yuan, Hongli
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1320-1332
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    • 2016
  • Light hydrocarbons accumulated in subsurface soil by long-term microseepage could favor the anomalous growth of indigenous hydrocarbon-oxidizing microorganisms, which could be crucial indicators of underlying petroleum reservoirs. Here, Illumina MiSeq sequencing of the 16S rRNA gene was conducted to determine the bacterial community structures in soil samples collected from three typical oil and gas fields at different locations in China. Incubation with n-butane at the laboratory scale was performed to confirm the presence of "universal microbes" in light-hydrocarbon microseepage ecosystems. The results indicated significantly higher bacterial diversity in next-to-well samples compared with background samples at two of the three sites, which were notably different to oil-contaminated environments. Variation partitioning analysis showed that the bacterial community structures above the oil and gas fields at the scale of the present study were shaped mainly by environmental parameters, and geographic location was able to explain only 7.05% of the variation independently. The linear discriminant analysis effect size method revealed that the oil and gas fields significantly favored the growth of Mycobacterium, Flavobacterium, and Pseudomonas, as well as other related bacteria. The relative abundance of Mycobacterium and Pseudomonas increased notably after n-butane cultivation, which highlighted their potential as biomarkers of underlying oil deposits. This work contributes to a broader perspective on the bacterial community structures shaped by long-term light-hydrocarbon microseepage and proposes relatively universal indicators, providing an additional resource for the improvement of microbial prospecting of oil and gas.

First Report of Bacterial Spot Disease Caused by Pseudomonas capsici on Castor Bean in Korea (Pseudomonas capsici에 의한 아주까리 세균점무늬병의 국내 첫 보고)

  • Heeil Do;Seung Yeup Lee;Bang Wool Lee;Hyeonheui Ham;Mi-Hyun Lee;Young Kee Lee
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.440-444
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    • 2023
  • In August 2021, water-soaking symptoms of bacterial spot disease were observed on castor bean in a field in Gangseo District, Busan. Bacteria isolated from the lesion when cultured on tryptic soy agar appeared to be nonmucoid and pale green. To confirm whether the isolates were the causative agent of the spot disease, they were inoculated onto healthy castor bean plants. The same symptoms were observed on the inoculated tissue, and the bacteria were reisolated from the lesion. Furthermore, the isolates were consistent with the biochemical and physiological features of Pseudomonas capsici. Sequencing analysis using 16S rRNA and housekeeping genes (gyrB, rpoD) showed that the isolates shared a high sequence similarity with P. capsici. These results confirmed that the strains belonged to P. capsici. To our knowledge, this is the first report of bacterial spot disease caused by P. capsici on castor bean in Korea.

Production and Characterization of a Novel Protease from Bacillus sp. RRM1 Under Solid State Fermentation

  • Rajkumar, Renganathan;Ranishree, Jayappriyan Kothilmozhian;Ramasamy, Rengasamy
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.627-636
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    • 2011
  • A commercially important alkaline protease, produced by Bacillus sp. RRM1 isolated from the red seaweed Kappaphycus alvarezii (Doty) Doty ex Silva, was first recognized and characterized in the present study. Identification of the isolated bacterium was done using both biochemical characterization as well as 16S rRNA gene sequencing. The bacterial strain, Bacillus sp. RRM1, produced a high level of protease using easily available, inexpensive agricultural residues solid-state fermentation (SSF). Among them, wheat bran was found to be the best substrate. Influences of process parameters such as moistening agents, moisture level, temperature, inoculum concentration, and co-carbon and co-nitrogen sources on the fermentation were also evaluated. Under optimized conditions, maximum protease production (i.e., 2081 U/g) was obtained from wheat bran, which is about 2-fold greater than the initial conditions. The protease enzyme was stable over a temperature range of 30-$60^{\circ}C$ and pH 6-12, with maximum activity at $50^{\circ}C$ and pH 9.0. Whereas the metal ions $Na^+$, $Ca^{2+}$, and $K^+$ enhanced the activity of the enzyme, others such as $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, $Co^{2+}$, and $Zn^{2+}$ had rendered negative effects. The activity of the enzyme was inhibited by EDTA and enhanced by $Cu^{2+}$ ions, thus indicating the nature of the enzyme as a metalloprotease. The enzyme showed extreme stability and activity even in the presence of detergents, surfactants, and organic solvents. Moreover, the present findings opened new vistas in the utilization of wheat bran, a cheap, abundantly available, and effective waste as a substrate for SSF.

The Distribution of Indicator Microorganisms and Identification of Antibiotic Resistant Strains in Domestic Animal Feces (가축 분변 유래 지표미생물 분포 및 항생제 내성 균주의 동정)

  • Kim, Jong-Geu;Lee, Jang-Hoon;Kwon, Hyuk-Ku
    • Journal of Environmental Health Sciences
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    • v.37 no.4
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    • pp.289-297
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    • 2011
  • Objectives: To estimate the microbial contaminant load discharged from livestock farms, we randomly selected livestock farmers of cattle, swine, and fowl and collected bacterial strains from domestic animals' feces and compost samples. Recently, as multi-antibiotic-resistant bacteria and super bacteria showing resistance to a variety of antibiotics have been reported one after another, the ecological and health hazard of antibiotic-resistant bacteria is emerging as an important issue. Methods: Monitored indicator microorganism constituents were totak coliform (TC), fecal coliform (FC), and aerobic bacteria. The multi-antibiotic-resistant bacteria were identified from investigated indicator microorganisms by 16S rRNA sequencing. Results: By microbiological analysis, the largest population of aerobic bacteria ($1.5{\times}10^5$ CFU/g) was found in cattle fecal compost, and total coliforms ($1.1{\times}10^7$ CFU/g) and fecal coliforms ($1.0{\times}10^5$ CFU/g) were found primarily in swine fecal compost, while the lowest population was found in fowl fecal compost. Among the 67 strains separated from aerobic bacteria, five strains expressing high antibiotic resistance were selected in each sample. We found the multi-antibiotic resistant strains to be Shigella boydii, Staphylococcus lentus, Acinetobacter sp. and Brevibacterium luteolum. Conclusions: These results suggest that increasing numbers of multi-antibiotic-resistant bacteria in the environment have a close relation to the reckless use of antibiotics with livestock.

Carbohydrate Fermentation Character of Bifidobacteria and Lactobacilli isolated from Feces of the Adult Women supplied with Goat Milk (산양유를 섭취한 성인 여성의 분변에서 분리한 Bifidobacteria와 Lactobacilli의 탄수화물 발효특성)

  • Choi, Suk-Ho;Lim, Young-soon;Ham, Jun-Sang;Jeong, Seok-Geun;Lee, Seung-Bae
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.103-110
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    • 2015
  • The objective of this study was to investigate the effects of consuming goat milk on the bacterial counts (colony forming units [CFU]) in adult women and to evaluate the carbohydrate fermentation capacity of bifidobacteria and lactobacilli isolated from their feces. Adult women who consumed goat milk (treatment group) had relatively higher CFU of bifidobacteria than did the control group, and the difference was significant (p<0.05) after 8 weeks. In total, 13 strains isolated from the feces of women in the treatment group were identified using 16S rRNA sequencing as Bifidobacterium adolescentis, B. longum, B. pseudocatenulatum, B. dentium, and Lactobacillus sakei. Similarly, 12 strains isolated from the feces of women in the control group included B. adolescentis, B. longum, L. ruminis, L. sakei, and B. pseudocatenulatum. All isolated bifidobacteria and lactobacilli fermented goat milk oligosaccharide and lactulose. All 7 strains of B. adolescentis fermented fructooligosaccharides, and 3 of the 4 B. pseudocatenulatum strains, 2 of the 3 L. sakei strains, and 1 of the 7 B. longum strains fermented fructooligosaccharides.

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