• Title/Summary/Keyword: bacterial isolate

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Anti-oxidative and Anti-bacterial Constituents from the Extracts of Rhododendron weyrichii Leaves (참꽃나무 잎 추출물 유래 항산화 및 항균 활성 성분)

  • Kim, Jung Eun;Jo, Seong Mi;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.4
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    • pp.341-351
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    • 2019
  • In this study, we investigated anti-oxidative and anti-bacterial constituents from Rhododendron weyrichii leaves. DPPH and ABTS+ radical scavenging activities were screened for the ethanol extract and solvent fractions. Potent scavenging activities were appeared from the extract, ethyl acetate (EtOAc) and n-butanol (BuOH) fractions. Upon the anti-bacterial tests using Staphylococcus epidermidis and Propionibacterium acnes, extract, n-hexane (Hex) and EtOAc fractions showed strong activities. To isolate the active constituents, the EtOAc fraction was further purified to afford five phytochemicals; ursolic acid (1), corosolic acid (2), asiatic acid (3), astragalin (4) and isoquercetin (5). All of the compounds 1-5 were isolated for the first time from this plant. Among the isolates, the compound 4 and 5 showed strong DPPH and ABTS+ radical scavenging activities. Also, compound 3 exhibited the most potent anti-bacterial activity. In addition, the content of astragalin isolated from this plant was determined by UPLC and the quantity was about 8.1 mg/g for the 70% ethanol extract and 34.8 mg/g for the EtOAc fraction. Based on these results, it is concluded that R. weyrichii extract could be potentially applicable as anti-oxidative and anti-bacterial ingredients in cosmetic formulations.

Isolation and Characterization of an Acyclic Isoprenoid from Semecarpus anacardium Linn. and its Antibacterial Potential in vitro - Antimicrobial Activity of Semecarpus anacardium Linn. Seeds -

  • Purushothaman, Ayyakkannu;Meenatchi, Packirisamy;Saravanan, Nallappan;Karuppaiah, Muthu;Sundaram, Ramalingam
    • Journal of Pharmacopuncture
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    • v.20 no.2
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    • pp.119-126
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    • 2017
  • Objectives: Semecarpus anacardium Linn. is a plant well-known for its antimicrobial, antidiabetic and anti-arthritic properties in the Ayurvedic and Siddha system of medicine. This has prompted the screening of this plant for antibacterial activity. The main aims of this study were to isolate compounds from the plant's seeds and to evaluate their antibacterial effects on clinical bacterial test strains. Methods: The n-butanolic concentrate of the seed extract was subjected to thin layer chromatography (TLC) and repeated silica gel column chromatography followed by elution with various solvents. The compound was identified based on observed spectral (IR, $^1H$ NMR, $^{13}C$ NMR and high-resolution mass spectrometry) data. The well diffusion method was employed to evaluate the antibacterial activities of the isolated acyclic isoprenoid compound (final concentration: $5-15{\mu}g/mL$) on four test bacterial strains, namely, Staphylococcus aureus (MTCC 96), Bacillus cereus (MTCC 430), Escherichia coli (MTCC 1689) and Acinetobacter baumannii (MTCC 9829). Results: Extensive spectroscopic studies showed the structure of the isolated compound to be an acyclic isoprenoid ($C_{21}H_{32}O$). Moreover, the isoprenoid showed a remarkable inhibition of bacterial growth at a concentration of $15{\mu}g/mL$ compared to the two other doses tested (5 and $10{\mu}g/mL$) and to tetracycline, a commercially available antibiotic that was used as a reference drug. Conclusion: The isolation of an antimicrobial compound from Semecarpus anacardium seeds validates the use of this plant in the treatment of infections. The isolated compound found to be active in this study could be useful for the development of new antimicrobial drugs.

Comparison of PFGE, IS6110-RFLP, and 24-Locus MIRU-VNTR for Molecular Epidemiologic Typing of Mycobacterium tuberculosis Isolates with Known Epidemic Connections

  • Jeon, Semi;Lim, Nara;Park, Sanghee;Park, Misun;Kim, Seonghan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.338-346
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    • 2018
  • Two molecular epidemiologic methods, IS6110 restriction fragment length polymorphism (IS6110-RFLP) and 24-locus mycobacterial interspersed repetitive unit-variable-number tandem repeat (MIRU-VNTR), are used worldwide in studies of Mycobacterium tuberculosis (MTB). Conversely, because of its poor resolution, pulsed-field gel electrophoresis (PFGE) is not widely used for MTB. In this study, we improved the 24-locus MIRU-VNTR and PFGE protocols and compared the effectiveness of these approaches for the molecular typing of MTB using 75 clinical isolates obtained from a cohort investigation of high-risk populations infected with MTB. The 24-locus MIRU-VNTR method demonstrated superior discriminatory ability, followed by PFGE and IS6110-RFLP. Next, we analyzed six isolates with clear epidemiologic connections; that is, isolates from patients who attended the same school. IS6110-RFLP and PFGE identified these samples as the same type. By contrast, according to MIRU-VNTR, two isolates differed from four other isolates at one locus each; one isolate was identified as Mtub29 and the other as QUB-26. In summary, the 24-locus MIRU-VNTR assay was the most useful molecular typing method among the three methods investigated due to its discriminatory power, short time required, and availability as an epidemiologic investigation tool. PFGE was the second-best method. Compared with the other loci assessed in the 24-locus MIRU-VNTR assay, the Mtub29 and QUB-26 loci appeared to exhibit greater variability during transmission.

Multiple Tolerances and Dye Decolorization Ability of a Novel Laccase Identified from Staphylococcus Haemolyticus

  • Li, Xingxing;Liu, Dongliang;Wu, Zhaowei;Li, Dan;Cai, Yifei;Lu, Yao;Zhao, Xin;Xue, Huping
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.615-621
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    • 2020
  • Laccases are multicopper oxidases with important industrial value. In the study, a novel laccase gene (mco) in a Staphylococcus haemolyticus isolate is identified and heterologously expressed in Escherichia coli. Mco shares less than 40% of amino acid sequence identities with the other characterized laccases, exhibiting the maximal activity at pH 4.0 and 60℃ with 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) as a substrate. Additionally, the Mco is tolerant to a wide range of pH, heavy metal ions and many organic solvents, and it has a high decolorization capability toward textile dyes in the absence of redox mediators. The characteristics of the Mco make this laccase potentially useful for industrial applications such as textile finishing. Based on BLASTN results, mco is found to be widely distributed in both the bacterial genome and bacterial plasmids. Its potential role in oxidative defense ability of staphylococci may contribute to the bacterial colonization and survival.

Isolation, Screening and Identification of Swine Gut Microbiota with Ochratoxin A Biodegradation Ability

  • Upadhaya, Santi Devi;Song, Jae-Yong;Park, Min-Ah;Seo, Ja-Kyeom;Yang, Liu;Lee, Chan-Ho;Cho, Kyung-J.;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.114-121
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    • 2012
  • The potential for ochratoxin A (OTA) degradation by swine intestinal microbiota was assessed in the current study. Intestinal content that was collected aseptically from swine was spiked with 100 ppb OTA and incubated for 6 and 12 h at $39^{\circ}C$. An OTA assay was conducted using the incubated samples, and it was found that 20% of the OTA toxin was detoxified, indicating the presence of microbes capable of OTA degradation. Twenty-eight bacterial species were isolated anaerobically in M 98-5 media and 45 bacterial species were isolated using nutrient broth aerobically. Screening results showed that one anaerobic bacterial isolate, named MM11, detoxified more than 75% of OTA in liquid media. Furthermore, 1.0 ppm OTA was degraded completely after 24 h incubation on a solid 'corn' substrate. The bacterium was identified by 16S rDNA sequencing as having 97% sequence similarity with Eubacterium biforme. The isolation of an OTA-degrading bacterium from the swine natural flora is of great importance for OTA biodegradation and may be a valuable potential source for OTA-degradation enzymes in industrial applications.

A report of 34 unrecorded bacterial species in Korea, belonging to the Actinobacteria

  • Ko, Kwan Su;Cha, Chang-Jun;Im, Wan-Taek;Kim, Seung-Bum;Seong, Chi-Nam;Bae, Jin-Woo;Jahng, Kwangyeop;Cho, Jang-Cheon;Joh, Ki-seong;Lee, Soon Dong
    • Journal of Species Research
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    • v.6 no.1
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    • pp.1-14
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    • 2017
  • As a subset study to discover indigenous prokaryotic species in Korea in 2014, a total of 34 bacterial strains assigned to the phylum Actinobacteria were isolated from various environmental samples collected from activate sludge, biotite, freshwater, gut of marine organisms, mud flat, sediment, soil, spent mushroom compost and sea water. On the basis of high 16S rRNA gene sequence similarity and a tight phylogenetic association with the closest species, it was revealed that each strain was assigned to independent and previously described bacterial species, with the exception of one isolate. There is no official report that these 34 species included in the phylum Actinobacteria have been described in Korea: 6 species of 5 genera in the order Corynebacteriales, 1 species of 1 genus in the order Frankiales, 2 species of 2 genera in the Micromonosporales, 14 species of 10 genera in Micrococcales, 2 species of 2 genera in the Propionibacteriales, 1 species of 1 genus in the Pseudonocardiales, 4 species of 2 genera in the Streptomycetales, 2 species of 2 genera in the Streptosporangiales and 1 species of 1 genus in the Solirubrobacterales. Gram reaction, cell and colony morphology, pigmentation, physiological characteristics, isolation sources and strain IDs are described in the section of species description.

Culture-Independent Analysis of Microbial Succession During Composting of Swine Slurry and Mushroom Cultural Wastes

  • Cho, Kye-Man;Lee, Sun-Mi;Math, Renukaradhya K.;Islam, Shah Md. Asraful;Kambiranda, Devaiah M.;Kim, Jong-Min;Yun, Myoung-Geun;Cho, Ji-Joong;Kim, Jong-Ok;Lee, Young-Han;Kim, Hoon;Yun, Han-Dae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1874-1883
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    • 2008
  • Bacterial diversity and the composition of individual communities during the composting process of swine and mushroom cultural wastes in a field-scale composter (Hazaka system) were examined using a PCR-based approach. The composting process was divided into six stages based on recorded temperature changes. Phylogenetic analysis of eighty 16S rRNA sequences from uncultured composting bacterial groups revealed the presence of representatives from three divisions, including plant pathogenic bacteria, high-molecule-degrading bacteria and spore-forming bacteria. The plant pathogen A. tumefaciens gradually decreased in abundance during the composting process and eventually disappeared during the thermophilic and cooling stage. A bacterium homologous to Bacillus humi first appeared at the early thermophilic stage and was established at the intermediate thermophilic, post-thermophilic, and cooling stages. It was not possible to isolate the B. humi during any of the stages using general culture techniques.

Isolation and identification of the abundant bacteria in dental caries in children (소아의 치아우식 부위별 우점 세균 분리 및 동정)

  • Kim, Eun-Mi
    • Journal of Korean society of Dental Hygiene
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    • v.18 no.5
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    • pp.843-852
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    • 2018
  • Objectives: The study aimed to isolate the abundant bacteria in dental caries in children and to investigate the bacterial species involved in addition to those that have been previously reported. Methods: The specimens were collected from the supragingival plaques of each dental caries area, pit and fissure caries, deep dentinal caries, smooth surface caries, and dental caries, and from healthy subjects in the control group. Bacteria were cultured from these specimens, DNA was extracted from the isolated bacteria, and the 16S rRNA gene sequences were analyzed and identified. Results: Based on the results of the 16S rRNA gene sequence analysis for the 90 strains of dominant bacteria from the 45 specimens, 5, 7, 8, 7, and 13 species were identified from the supragingival plaques from healthy teeth, pit and fissure caries, deep dentinal caries, smooth surface caries, and dental caries, respectively. In healthy teeth, Actinomyces naeslundii dominated. Corynebacterium durum, Ralstonia pickettii, and Streptococcus intermedius showed equal distribution. The dominant bacterial species in dental caries, S. sanguinis, showed the greatest difference in prevalence in pit and fissure caries. In deep dentinal caries, S. mutans and Lactobacillus rhamnosus were dominant; in smooth surface caries, S. mutans and S. sanguinis were dominant; and in the supragingival plaques of dental caries, S. sanguinis and S. mutans were dominant. Conclusions: The bacterial species isolated from dental caries encompassed four phyla, eight genera, and 22 species. In addition, the SS1-2 strain, belonging to the genus Neisseria, was identified as a new species from among the isolated strains.

Lipooligosaccharide biosynthesis genes of nontypable Haemophilus influenzae 2019

  • Lee, Na-Gyong;Melvin G. Sunshine;Jeffery J. Engstrom;Bradford W. Gibson;Michael A. Apicella
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.11a
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    • pp.65-73
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    • 1996
  • LPS/LOS, the compound found only in gram-negative bacterial outer membrane, plays important roles in bacterial maintenance as well as its pathogenesis. We isolated and characterized several genes required for NTHi 2019 LOS biosynthesis, which encode enzymes required for sugar substrate synthesis or the transfer of substrates to receptor molecules. The htrB gene, however, appears to have more complex role. It has acryltransferase activity as well as various other activity, which may control regulation of LOS biosynthesis as well as its pathogenicity. Evidences supporting the latter come from the observations that the lipid A of the B29 induced significantly less TNF ${\alpha}$ from macrophages than that of the wild type LOS (unpublished data). H. influenzae A2-htrB mutant strain was also significantly less invasive than the wild type strain. The structural similarities of the enterobacterial LPS and the Haemophilus LOS enabled us to isolate the NTHi 2019 genes involved in LOS biosynthesis genes by using the S. typhimurium LPS deep core mutants. While a similar approach has been used for E. coli, this technique for selection of an LPS phenotype has not been applied to nonenterobacterial species. The difficulties inherent in the molecular manipulation of organism such as Neisseria and Haemophilus species make this approach particularly attractive in the identification and cloning LOS genes. Studies on genetic features of LPS/LOS biosynthesis would be useful for understanding bacterial pathogenesis as well as for developing vaccines for these gram-negative pathogenic bacteria.

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Effects of Heavy Metal and pH on Bacterial Growth Isolated from the Contaminated Smelter Soil (제련소 인근 토양에서 분리한 박테리아 생장에 미치는 중금속 및 pH 영향)

  • Keum, Mi-Jung;Yoon, Min-Ho;Nam, In-Hyun
    • Journal of Soil and Groundwater Environment
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    • v.20 no.4
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    • pp.113-121
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    • 2015
  • The contaminated soil at abandoned smelter areas present challenge for remediation, as the degraded materials are typically deficient in nutrients, and rich in toxic heavy metals and metalloids. Bioremediation technique is to isolate new strains of microorganisms and develop successful protocols for reducing metal toxicity with heavy metal tolerant species. The present study collected metal contaminated soil and characterized for pH and EC values, and heavy metal contents. The pH value was 5.80, representing slightly acidic soil, and EC value was 13.47 mS/m. ICP-AES analytical results showed that the collected soil samples were highly contaminated with various heavy metals and metalloids such as lead (183.0 mg/kg), copper (98.6 mg/kg), zinc (91.6 mg/kg), and arsenic (48.1 mg/kg), respectively. In this study, a bacterial strain, Bacillus cereus KM-15, capable of adsorbing the heavy metals was isolated from the contaminated soils by selective enrichment and characterized to apply for the bioremediation. The effects of heavy metal on the growth of the Bacillus cereus KM-15 was determined in liquid cultures. The results showed that 100 mg/L arsenic, lead, and zinc did not affect the growth of KM-15, while the bacterial growth was strongly inhibited by copper at the same concentration. Further, the ability of the bacteria to adsorb heavy metals was evaluated.