• Title/Summary/Keyword: Agar diffusion assay

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Antibacterial activity of isolated bacteria against Propionibacterium acnes causing acne vulgaris (여드름을 유발하는 Propionibacterium acnes에 대한 분리 세균들의 항균활성)

  • Lee, Da-Sol;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.272-279
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    • 2018
  • This study was carried out to evaluate antimicrobial activity of isolated bacteria from various soils against two strains of Propionibacterium acnes causing acne vulgaris. Among several hundreds of bacterial strains, Paenibacillus elgii DS381, Paenibacillus elgii DS1515, Burkholderia gladioli DS518, and Streptomyces lienomycini DS620 showed high antimicrobial activities against the strains of P. acnes. All isolated bacteria showed 15.5 to 34.3 mm inhibition zone diameter in an agar well diffusion test, and especially DS620 showed the highest inhibition zone diameters (28.3~34.3 mm). Antibacterial substances were expected as lipopeptide (pelgipeptin and paenipeptin) from strains DS381 and DS1515, protease from DS518, and anthracycline antibiotic (daunomycinone) from DS620, and all these showed very low minimum inhibitory concentration [DS381 and DS1515 (0.078 mg/ml), DS518 (0.312 mg/ml), DS620 (0.000078 mg/ml)] against P. acnes. These antibacterial substances could completely kill P. acnes within 24 h in a time-kill assay. These results suggest that antibacterial substances produced by these bacteria may be utilized as useful antimicrobial agent against P. acnes and treatment medicine for acne vulgaris.

Expression of Antimicrobial Peptide (AMP), Moricin Using SUMO Fusion Tag in Escherichia coli (대장균에서 SUMO fusion tag을 이용하여 항균펩타이드인 moricin의 발현)

  • Ahn, Dong-gyu;Park, Sun Ill;Kim, Soon Young
    • Journal of Life Science
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    • v.32 no.12
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    • pp.956-961
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    • 2022
  • Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.

Construction of a Plant Chloroplast Transformation Vector to Produce the Antimicrobial Peptide Stomoxyn (대장균에서 항균 펩타이드 stomoxyn을 생산하기 위한 형질전환 벡터 제작)

  • Jin Gyu Go;Hyeon Ho Gil;Soon Young Kim
    • Journal of Life Science
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    • v.34 no.7
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    • pp.493-499
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    • 2024
  • Antimicrobial peptides are antimicrobial substances inherent in animals and plants, with strong antibacterial activity even in small amounts and with various other functions such as antiviral and antioxidant actions. Plants can be grown with just water and sunlight, allowing for their mass production at low costs. However, transforming a chloroplast into one that produces antimicrobial peptides, rather than growing plants, increases the amount of protein expression and minimizes contamination of the ecosystem because gene transfer by pollen does not occur. In that context, using transgenic plant chloroplasts to produce recombinant proteins increases protein degradation and reduces the solubility of proteins. To solve this problem, we fused SUMO, a fusion protein, with a recombinant protein. We also used a 6xHis tag to purify the fusion protein. The antimicrobial peptide stomoxyn is an antibacterial substance found in stable flies. Stomoxyn has an α-helix structure and is amphiphilic, which allows it to dissolve bacterial cell membranes. In this study, we constructed a transformation vector to express stomoxyn in both plant chloroplasts and Escherichia coli and used this vector to confirm the expression of stomoxyn in E. coli. The expression of the protein was then confirmed in E. coli using a transformation vector. The expressed stomoxyn was purified by nickel column and SUMOase treatment, and its antibacterial activity was confirmed using an agar diffusion assay. The EGFP gene was used to ensure that the transformed vector was inserted into the chloroplast.

In Vitro Antagonistic Activity Evaluation of Lactic Acid Bacteria (LAB) Combined with Cellulase Enzyme Against Campylobacter jejuni Growth in Co-Culture

  • Dubois-Dauphin, Robin;Sabrina, Vandeplas;Isabelle, Didderen;Christopher, Marcq;Andre, Thewis;Philippe, Thonart
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.62-70
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    • 2011
  • The antibacterial effects of nine lactic acid bacteria (LAB) against Campylobacter jejuni were investigated by using agar gel diffusion and co-culture assays. Some differences were recorded between the inhibition effects measured with these two methods. Only two LAB, Lb. pentosus CWBI B78 and E. faecium THT, exhibited a clear anti- Campylobacter activity in co-culture assay with dehydrated poultry excreta mixed with ground straw (DPE/GS) as the only growth substrate source. It was observed that the supplementation of such medium with a cellulase A complex (Beldem S.A.) enhanced the antimicrobial effect of both LAB strains. The co-culture medium acidification and the C. jejuni were positively correlated with the cellulase A concentration. The antibacterial effect was characterized by the lactic acid production from the homofermentative E. faecium THT and the lactic and acetic acids production from the heterofermentative Lb. pentosus CWBI B78. The antagonistic properties of LAB strains and enzyme combination could be used in strategies aiming at the reduction of Campylobacter prevalence in the poultry production chain and consequently the risk of human infection.

Characterization and antimicrobial efficacy of Portland cement impregnated with silver nanoparticles

  • Nam, Ki Young
    • The Journal of Advanced Prosthodontics
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    • v.9 no.3
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    • pp.217-223
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    • 2017
  • PURPOSE. This study investigated the effects of silver nanoparticle (SN) loading into hydraulic calcium silicate-based Portland cement on its mechanical, antibacterial behavior and biocompatibility as a novel dental bone substitute. MATERIALS AND METHODS. Chemically reduced colloidal SN were combined with Portland cement (PC) by the concentrations of 0 (control), 1.0, 3.0, and 5.0 wt%. The physico-mechanical properties of silver-Portland cement nanocomposites (SPNC) were investigated through X-ray diffraction (XRD), setting time, compressive strength, solubility, and silver ion elution. Antimicrobial properties of SPNC were tested by agar diffusion against Streptococcus mutans and Streptococcus sobrinus. Cytotoxic evaluation for human gingival fibroblast (HGF) was performed by MTS assay. RESULTS. XRD certified that SN was successfully impregnated in PC. SPNC at above 3.0 wt% significantly reduced both initial and final setting times compared to control PC. No statistical differences of the compressive strength values were detected after SN loadings, and solubility rates of SPNC were below 3.0%, which are acceptable by ADA guidelines. Ag ion elutions from SPNC were confirmed with dose-dependence on the concentrations of SN added. SPNC of 5.0 wt% inhibited the growth of Streptococci, whereas no antimicrobial activity was shown in control PC. SPNC revealed no cytotoxic effects to HGF following ISO 10993 (cell viability > 70%). CONCLUSION. Addition of SN promoted the antibacterial activity and favored the bio-mechanical properties of PC; thus, SPNC could be a candidate for the futuristic dental biomaterial. For clinical warrant, further studies including the inhibitory mechanism, in vivo and long-term researches are still required.

Antibacterial Activity and Probiotic Properties of Lactic Acid Bacteria from Korean Intestine Origin (한국인 장관에서 분리한 유산균의 항균활성 및 프로바이오틱스 특성 연구)

  • Kang, Chang-Ho;Kim, Yong Gyeong;Han, Seul Hwa;Jeong, Yulah;Paek, Nam-Soo
    • KSBB Journal
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    • v.32 no.2
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    • pp.153-159
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    • 2017
  • The purpose of this study was to investigate the probiotic properties of lactic acid bacterial strains isolated from human feces. The properties were tested on the basis of guideline for probiotic selection protocol such as tolerance for acid or bile salt, autoaggregation, antibiotic resistance, and antimicrobial activity. Total 25 lactic acid bacteria were isolated from human feces, and their antibacterial activity was tested against Staphylococcus aureus, Escherichia coli, E. coli O157:H7, Vibrio parahaemolyticus, V. alginolyticus using an agar diffusion assay. Among them, 4 selected strains were identified by analysis of their 16S rRNA, as Lactobacillus rhamnosus MG316, L. acidophilus MG501, L. reuteri MG505, and L. gasseri MG570. Results show that resistance to low pH and bile salts. Also, the selected strains were resistant to bile acid up to 3% and their autoaggregation rates were as high as 60%. All strains tested were resistance to nalidixic acid and kanamycin.

Antibacterial effect of bamboo charcoal on Streptococcus mutans (S. mutans에 대한 대나무 숯의 항균력에 관한 연구)

  • Choi, Mi-Suk;Ahn, Kwon-Suk
    • Journal of Korean society of Dental Hygiene
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    • v.14 no.1
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    • pp.95-100
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    • 2014
  • Objectives : The purpose of the present study was to investigate the effect of bamboo charcoal on Streptococcus mutans which is one of the most important causative agents of dental caries. Methods : S. mutans was incubated with or without bamboo charcoal and then changes were observed in its cell viability and antibacterial effect. Oral epithelial cells viabillity(human gingival fibroblast, HGF) was performed using MTT assay. Antibacterial effect was analyzed using a dilution plating method and agar diffusion method. Results : Oral epithelial cells, human gingival fibroblast (HGF) showed a tendency to increase in bamboo charcoal treatment solution concentrations(0.5, 1, 2, 3, 5, 10%). The bamboo charcoal had an antibacterial effect on S. mutans. Antibacterial effect of bamboo charcoal for the bacterium was 58%. Charcoal concentration of 2% and 5% in the inhibition zone showed a minimal growth, but the concentration of 10% bamboo charcoal in inhibition zone revealed a conspicuous antibacterial activity. Conclusions : Overall results suggested that the bamboo charcoal proved to be bactericidal effect on S. mutans.

The Antimicrobial Activity of Essential Oil from Dracocephalum foetidum against Pathogenic Microorganisms

  • Lee, Saet-Byoul;Cha, Kwang-Hyun;Kim, Su-Nam;Altantsetseg, Shataryn;Shatar, Sanduin;Sarangerel, Oidovsambuu;Nho, Chu-Won
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.53-57
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    • 2007
  • A number of essential oils from Mongolian aromatic plants are claimed to have antimicrobial activities. The essential oil of Dracocephalum foetidum, a popular essential oil used in Mongolian traditional medicine, was examined for its antimicrobial activity. Eight human pathogenic microorganisms including B. subtilis, S. aureus, M. lutens, E. hirae, S. mutans, E. coli, C. albicans, and S. cerevisiae were examined. The essential oil of Dracocephalum foetidum exhibited strong antimicrobial activity against most of the pathogenic bacteria and yeast strains that were tested; by both the agar diffusion method and the minimum inhibitory concentration (MIC) assay ($MIC\;range\;was\;26-2592{\mu}g/ml$). Interestingly, Dracocephalum foetidum even showed antimicrobial activity against methicilin-resistant Staphylococcus aureus (MRSA) strains. We also analyzed the chemical composition of the oil by GC-MS and identified several major components, including n-Mentha-1,8-dien-10-al, limonene, geranial, and neral.

Drug adsorption and anti-microbial activity of functionalized multiwalled carbon nanotubes

  • Saxena, Megha;Mittal, Disha;Boudh, Richa;Kumar, Kapinder;Verma, Anita K.;Saxena, Reena
    • Advances in nano research
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    • v.11 no.6
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    • pp.667-678
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    • 2021
  • Multiwalled carbon nanotubes (MWCNTs) were first oxidized (O-CNTs) to introduce carboxylic group and then further functionalized (F-CNTs) with m-phenylenediamine, which was confirmed by FTIR and SEM. It was used as an effective adsorbent for the adsorptive removal of diclofenac drug from water. Under optimum conditions of pH 6, stirring speed 600 rpm, the maximum adsorption capacity obtained was 532 mg g-1 which is superior to the values reported in literature. The adsorption was quite rapid as 25 mg L-1 drug solution was adsorbed in only 3 minutes of contact time with 10 mg of adsorbent dose. The adsorption kinetics and isotherms were studied using various models to evaluate the adsorption process. The results showed that the data best fit in kinetics pseudo-second order and Langmuir isotherm model. Furthermore, the oxidized and functionalized MWCNTs were applied on gram-negative Escherichia coli and gram-positive Staphylococcus aureus using agar disc diffusion assay to validate their anti-microbial activity. Results were unique as both oxidized and functionalized MWCNTs were equally active against both E. coli and S. aureus. The newly synthesized F-CNTs have great potential in water treatment, with their dual action of removing drug and pathogens from water, makes it potential applicant to save environment.

Inhibition of Quorum Sensing Regulated Virulence Factors and Biofilm Formation by Eucalyptus globulus against Multidrug-Resistant Pseudomonas aeruginosa

  • Sagar, Pankaj Kumar;Sharma, Poonam;Singh, Rambir
    • Journal of Pharmacopuncture
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    • v.25 no.1
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    • pp.37-45
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    • 2022
  • Objectives: The quorum-sensing-inhibitory and anti-biofilm activities of the methanol extract of E. globulus leaves were determined against clinically isolated multidrug-resistant Pseudomonas aeruginosa. Methods: The preliminary anti-quorum-sensing (AQS) activity of eucalyptus was investigated against a biosensor strain Chromobacterium violaceum ATCC 12472 (CV12472) by using the agar well diffusion method. The effect of sub-minimum inhibitory concentrations (sub-MICs) of the methanol extract of eucalyptus on different quorum-sensing-regulated virulence factors, such as swarming motility, pyocyanin pigment, exopolysaccharide (EPS), and biofilm formation, against clinical isolates (CIs 2, 3, and 4) and reference PA01 of Pseudomonas aeruginosa were determined using the swarm diameter (mm)-measurement method, chloroform extraction method, phenol (5%)-sulphuric acid (concentrated) method, and the microtiter plate assay respectively, and the inhibition (%) in formation were calculated. Results: The preliminary AQS activity (violacein pigment inhibition) of eucalyptus was confirmed against Chromobacterium violaceum ATCC 12472 (CV12472). The eucalyptus extract also showed concentration-dependent inhibition (%) of swarming motility, pyocyanin pigment, EPS, and biofilm formation in different CIs and PA01 of P. aeruginosa. Conclusion: Our results revealed the effectiveness of the E. globulus extract for the regulation of quorum-sensing-dependent virulence factors and biofilm formation at a reduced dose (sub-MICs) and suggest that E. globulus may be a therapeutic agent for curing and controlling bacterial infection and thereby reducing the possibility of resistance development in pathogenic strains.